BIOCHEMISTRY

SHIFT 3
Blood Generalities 17 JAN 2023 01
Dr. Judelyn T. Uy AY 2022-2023

TABLE OF CONTENTS 1. BLOOD GENERALITIES AND RBC METABOLISM (DOK)

1. Blood Generalities and RBC Metabolism ….………….1 1.1. MAJOR FUNCTIONS OF BLOOD
1.1. Medical Importance
2. Four Main Components…………………………….……..2
• Primary function to transport molecules around
2.1. Physical Characteristics of Blood
2.2. Composition of Blood the body to support critical metabolic processes
2.3. Hematopoietic Stem Cells
2.4. Cytokines
2.5. JAK-STAT Pathway
2.6. Hematopoiesis
3. Red Blood Cells (Erythrocytes)....................................6
3.1. Regulation of RBC Production
3.2. Erythropoiesis
3.3. Red Blood Cells (Erythrocytes)
3.4. RBC Membrane
3.5. Aged Red Blood Cells
4. Red Blood Cell Metabolism…………………………..…12 Fig. 1.1. Transport Function of Blood
4.1. Embden-Meyerhof Pathway
4.2. Rapoport-Luebering Shunt • Facilitates delivery of absorbed nutrients to
4.3. Hexose Monophosphate Shunt different organs of the body
4.4. Methemoglobin Reduction Pathway
4.5. Carbonic Anhydrase • Transport many hormones to its target organ
4.6. Biochemical Basis of ABO Blood Group System or tissue for regulation of metabolic
homeostasis
TRANSPORT
5. Disorders affecting RBC……………………….……….22
6. Platelets (Thrombocytes)............................................22 • Removes metabolic waste products either
6.1. Function of Platelets (Thrombocytes) through the skin, kidneys, or intestines
6.2. Major Metabolic Pathways • Examples:
6.3. Disorders affecting Platelets o Respiratory system: transport process
7. White Blood Cells (Leukocytes).................................24
during gas exchange
7.1. Main types of Leukocytes
7.2. Origin of Leukocytes o Delivers oxygen from lungs to other
7.3. General Characteristics of Leukocytes tissues and CO2 (travels mostly as
7.4. Neutrophils bicarbonate) is transported from
7.5. Eosinophils tissues as a waste product of cellular
7.6. Basophils respiration back to the lungs
7.7. Monocytes
7.8. Lymphocytes
7.9. Major Metabolic Pathways
7.10. Leukocyte Functions
7.11. Disorders affecting WBC
8. Plasma……………………………………………………29
9. Plasma Proteins…………………………………….…..29
9.1. Effect of Plasma Proteins in Osmotic Pressure
9.2. Plasma Proteins Overview
9.3. Albumin
9.4. Fibrinogen
9.5. Globulins
9.6. Plasma Proteins in DIagnosis of Diseases Fig. 1.1-2 Transport Process and Exchange
9.7. Plasma Protein Electrophoresis in Diagnosis of
Diseases

LEGEND
★ Take note / Important ☛ Prof verbatim
✎ Textbook info ✂ Previous/Other trans info

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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

• Regulate body temperature by redistributing • In the blood, there are WBCs and circulating
body's heat antibodies that help protect against
• Examples microorganisms and foreign substances
o Febrile: blood vessels vasodilate = • Inflammation occurs in the blood vessels due to
promote heat loss the release of inflammatory mediators
o Cold environment: blood vessels • Blood undergoes clotting in response to
vasoconstrict = keep core body vascular injury such as bleeding where a
temperature in control series of clotting and anti-clotting factors are
o Cold → Constrict kept in balance
o To make sure that clotting happens
only during vascular injury
o Circulating platelets would help protect
us from blood loss by forming a mesh
and plug to coagulate the blood and
stop the bleeding

PROTECTION
REGULATION

Fig. 1.1-3. Vascular regulation

• Buffers found in the blood (carbonic acid,

bicarbonate ion, carbon dioxide) need to
maintain normal acid-balance
• Capillaries are where exchange of fluid and
electrolytes occur between the circulating fluid
and the body tissues

Fig. 1.1-5. Clotting Process

2. FOUR MAIN COMPONENTS (DOK, MOO, MYG)

Fig. 1.1-4. Exchange of Electrolytes
• The myriad of functions is carried out by diverse
components of the blood including the red blood cells,
platelets, white blood cells, and plasma.

2.1. PHYSICAL CHARACTERISTICS OF BLOOD (DOK)

• Blood is a free-flowing fluid that is

Viscosity slightly denser and is 4.5-5.5x more
viscous than water
• Arterial – bright red
• Venous – dark red
• Human blood is characteristically a
red fluid where hemoglobin is the
principal determinant of blood color
Color attributed to the presence of the
heme protein
o When oxygenated, such
as in the arteries, the
oxygen binds with the
heme iron and appear
as bright red in color

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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

o Darkens when the iron (Instructor’s Slides)

is deoxygenated in the
venous blood

Fig. 2.2-2. Characteristics of Serum and Plasma

(Instructor’s Slides)
Fig. 2.1-1. Arterial and Venous Blood
(Instructor’s Slides) • Usually does not contain
anticoagulants
• Blood has slightly basic pH • When whole blood is centrifuged and
pH
• 7.35-7.45 left to clot, the liquid layer at the top
• 7-8% of total body weight Red top is serum
• Average blood volume varies with o Formed elements are in a
age and is relatively higher in infants fibrin clot
and neonates compared to adults o Serum does not contain any
• Males: 5-6L (Generally higher than clotting factor
in females) • Has added anticoagulants, which
• Females: 4-5L prevents clotting of the blood
• Healthy adults can lose almost 20% • When centrifuged, liquid portion is
Volume now called the plasma
of blood volume before symptoms of
dizziness or restlessness begin o Plasma is different from
Purple top
• 40% of volume before hypovolemic serum because clotting
shock sets in factors are still present in the
liquid portion of the plasma
• Conversely, higher than normal
and formed elements are
blood volume may also cause
easily separated for study
hypertension, heart failure, and
aneurysms
• 38°C normal temperature of blood
Temperature • Slightly higher than the 37 C of the
normal body temperature

2.2. COMPOSITION OF BLOOD (DOK, MOO)

• When a sample of whole blood is extracted and
centrifuged, there is separation of the liquid portion of
the blood at the top layer and the formed elements at
the bottom layer
• Liquid portion
o Serum or plasma depending on how the whole
blood sample was collected

Fig. 2.2-3. Composition of Purple Top Tube Blood Sample

(Instructor’s Slides)

• Aside from plasma, there are two more layers at the

bottom:
o Red Blood Cells: Thicker, reddish layer at the
lowermost portion of the test tube that
comprises mostly of red blood cells settling at
the bottom because they are the heaviest
elements in the blood
Fig. 2.2-1. Composition of Whole Blood

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o Buffy Coat: Thin, white, middle layer (fraction

of coagulated blood) containing the white blood
cells and platelets

Fig. 2.2-4c. Components of Whole Blood

(Instructor’s Slides)

Fig. 2.2-4a. Components of Whole Blood • Blood comprises 8% of total body weight
(Instructor’s Slides) o Pale, straw-colored fluid called plasma (55%)
▪ About 90% water, 7% proteins,
• In the circulation, whole blood is composed of plasma about 2% other solutes
and formed elements o Formed elements (45%)
o Serum is only formed during blood extraction ▪ >99% RBCs
when clotting factors, specifically fibrinogen, ▪ <1% WBCs and platelets
are removed from plasma • Collectively seen as white
layer in between plasma and
RBCs called buffy coat

✂ LECTURETTE KAHOOT (Composition of Blood)

What will be left from the blood if formed elements
and clotting factors were removed?
A. Red blood cells
B. Plasma
C. Platelets & WBC
D. Serum
Fig. 2.2-4b. Components of Whole Blood Answer: D
(Instructor’s Slides)
• Removal of formed elements and clotting factors
– serum
• Of the approximate 4-6 L of blood circulating in the
• Removal of formed elements only – plasma
body:
o Plasma constitutes about 55%, and is made up • Red top tube (no anticoagulants) – serum (fluid
of: portion) is collected after centrifugation
▪ Water (90%) • Tubes with anticoagulant (purple, blue, green,
▪ Solutes (10%) made up of: yellow) – plasma (fluid portion) is collected
• Organic compounds (6-8%) • Buffy coat - composed of WBC and platelets
which include lipids,
hormones, immunoglobulins, What comprises majority of solid particles in the
proteins, clotting factors and blood plasma?
fibrinogen A. Electrolytes
• Inorganic compounds (2- B. Proteins
4%) include salts, acids, or C. Clotting Factors
bases D. Nutrients
▪ Plasma fluid also continuously
suspends the formed elements and Answer: B
enables them to circulate throughout • 90% of plasma is composed of water while the
the body rest are the solid particles, 7-8% of which are the
o Formed elements (RBCs along with WBCs plasma proteins.
and platelets) make up about 45%, and is made
up of Which cellular component makes up the majority of
▪ Red blood cells (99%) the formed elements in the blood?
▪ Others (1%) A. Erythrocytes
• WBCs and platelets comprise B. Neutrophils
only <1% of the formed C. Lymphocytes
elements D. Platelets

Answer: A

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• More than 99% of the cellular component is
RBCs.
2.3. HEMATOPOIETIC STEM CELLS (MOO)
• All formed elements from blood have a common
precursor derived from hematopoietic stem cells
o These cells migrate from sites of
hematopoiesis to reside in the bone marrow
• In the presence of appropriate signals, hematopoietic
stem cells will undergo proliferation, differentiation, and
maturation
o These processes are regulated by a set of
secreted glycoproteins called cytokines
2.4. CYTOKINES (MOO)
• Subtype of growth factors secreted by specific cells of
hematopoietic and immune systems
• Cell-signaling molecules that regulate amplitude and
duration of the responses
• Mediate and regulate immunity, inflammation, and
hematopoiesis
• Examples:
o Interleukins (IL)
o Interferons (IFN)
o Chemokines
o Colony stimulating factors (CSF)
o Tumor necrosis factors (TNF)
✂ LECTURETTE KAHOOT (Cytokines)
Which cytokine attracts and activates migrating
leukocytes to a site of injury or infection?
1. Interleukins
2. Chemokines
3. Interferons
4. Tumor necrosis factor
Answer: B
• Chemotaxis would refer to migration of WBCs or
leukocytes to site of injury or infection, induced
by chemokines
• Interleukins, interferons, and TNF are some of
the cytokines that can activate proliferation and
differentiation of the different cell components
2.5. JAK STAT PATHWAY (MOO, MYG)
Fig 2.5-1. JAK-STAT Pathway
(Instructor’s Slides)
• Most of the cytokines are recognized by the receptors
which activate the Janus-Kinase – Signal Transducer
and Activator of Transcription (JAK-STAT) Pathway
BIOCHEMISTRY
• Janus Kinases
o Family of cytoplasmic tyrosine kinases that
links cytokine signaling from membrane
receptors to the STAT transcription factor
o Usually active when phosphorylated
Cytokine binding to receptors initiates
1 dimerization and activation of JAKs and favors
phosphorylation of JAK tyrosine residues
JAK dimerization and phosphorylation promotes
binding of additional signal transduction
2
molecules to the STAT family of transcription
factors
Binding and phosphorylation of STAT promotes
3 its dimerization and its translocation to the
nucleus, where it activates gene transcription
Activation of gene transcription would then
4 promote proliferation of specific cell lineages
from the hematopoietic stem cells
Silencer of Cytokine Signaling (SOCS)
proteins (4&5)
• Negative regulators of the JAK-STAT
5 pathway
• Either bind to the phosphorylated receptors
and inhibit further binding of STAT, or bind to
the JAKs itself and inhibit them
Specific cytokines bind to the receptors, activate
the JAK-STAT pathway, and initiate
6
transcription of a specific cell type or cell
lineage
✂ LECTURETTE KAHOOT (JAK-STAT)
This pathway is activated upon cytokine binding for
differentiation and proliferation of hematopoietic and
immune cells.
• Answer: Janus protein tyrosine kinase (JAK) – Signal
Transducer and Activator of Transcription (STAT)
pathway
Rationale:
• EPO will stimulate RBC synthesis -> binds to tyrosine
kinase (JAK) receptors within the bone marrow ->
phosphorylation of STAT -> promotes dimerization -
> gene expression/transcription
• In gene transcription, synthesis of proteins is
stimulated; there is activation of RBC lineage.
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

2.6. HEMATOPOIESIS (MYG, WRITER) • Production of all the cellular components of the
blood will then form three main types, the most
numerous of which are the red cells, followed by the
tiny platelets, and the least frequent cells, the white
blood cells

3. RED BLOOD CELLS (MYG, KSZ, 2PC)

ERYTHROCYTES
• Most abundant cellular component
Definition of blood (40-45% of blood); gives
blood its red color
• 4.7 - 6.1 x 106 /mm3 (uL) in males
RBC • 4.2 - 5.4 x 106 /mm3 (uL) in
count NV females
• Comparison: Females < Males
Fig 2.6-1. Hematopoiesis HEMATOCRIT LEVEL
(Instructor’s Slide)
• Volume of erythrocytes compared
to the total blood volume
• Definition
In the presence of appropriate signals, the body • Proportion of blood by volume
produces the different types of cells which undergo a consisting of RBC
series of differentiation, proliferation, and maturation • 45 - 52% in males
before they are released into the circulation. Hct level
• 37 - 48% in females
• Hematopoietic stem cell proliferation & differentiation NV
• Comparison: Females < Males
into common myeloid progenitor cells is stimulated by:
• Ratio of the volume of RBC to the
o Stem cell factor (SCF) and Granulocyte-
volume of other components in
monocyte colony-stimulating factor (GM-
whole blood
CSF) collaborate with the following:
Example • Ex: if there is 45% Hct, there is
▪ IL1
45mL of RBC in 100mL of blood
▪ IL3
▪ IL6 • Hct level is directly proportional
to the RBC count
• Myeloid progenitor cells then differentiate into:
HEMOGLOBIN LEVEL
• RBCs if directed by erythropoietin
• Platelets if directed by thrombopoietin • Represents the protein component
• Granulocytes (neutrophils, eosinophils, within each RBC
basophils) and monocytes if stimulated by: Definition • Delivers oxygen to the organs and
o Stem cell growth factor, particularly GM- tissues; carries CO2 from tissues to
CSF together with IL5 and IL6 the lungs
• Formation of lymphoid progenitor cells and maturation • 13.5 - 17.5 g/dL in males
HgB NV
into B lymphocytes (in the bone marrow) and T • 12.0 - 15.5 g/dL in females
lymphocytes (in the thymus) is stimulated by:
o Tumor necrosis factor (TNFα) ANEMIA
o Transforming growth factor β1 (TGFβ1) • Occurs when Hgb concentration is below normal
o IL2 values
o IL7 Red Cell Size
o IL12 • Normal Size - NORMOCYTIC
o FLT3 ligand • Small - MICROCYTIC
• Large - MACROCYTIC
• Measured by Mean Corpuscular
Volume (MCV)

Categories

• Normocytic anemia = 80-100 fL

• Microcytic anemia < 80 fL
• Macrocytic anemia > 100 fL
Fig 2.6-2. Formed Elements of Blood
(Instructor’s Slide) Hemoglobin Concentration

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• Normal Size - NORMOCHROMIC
• Small - HYPOCHROMIC
• Large - HYPERCHROMIC
• Measured by Mean Hemoglobin
Concentration (MCHC)
o Used to help identify the average
Hgb concentration within each
individual RBC
• MCHC NV: 32 – 36 g/dL
• If below NV, RBC is hypochromic
o Will appear lighter
• If above NV, RBC is hyperchromic
o Will appear darker
✂ LECTURETTE KAHOOT (Anemias)
What type of anemia present with low MCV and
normal MCHC values during complete blood count
(CBC) test
• Microcytic, normochromic
• Normocytic, hypochromic
• Microcytic, hypochromic
• Microcytic, hyperchromic
Answer: Microcytic, normochromic
Rationale:
• MCV or mean corpuscular volume will measure
the size so terms that end in -cytic
• MCHC or mean corpuscular hemoglobin content
measures color so terms that end in -chromic
3.1. REGULATION OF RBC PRODUCTION
• ☛ Nutrients are needed for proper RBC
production, and deficiency of which can lead to
anemia.
Nutrients
Iron
• Essential component of the
heme group in a hemoglobin
molecule
• 20% of dietary iron is
absorbed
o ☛ Less than 20% of the
iron consumed is absorbed
Definition
on average
• Heme iron from animal foods
(e.g., meat, poultry, and fish)
are better absorbed/absorbed
more efficiently compared to
non-heme iron from plant
foods
BIOCHEMISTRY
Fig. 3.1 -1. Iron Transport
• Ferroportin facilitates the
transport of iron across the
Iron Transport intestinal cell plasma
membrane
• Upon absorption, iron will be
bound to transferrin in the
circulation
• The bone marrow, liver, and
spleen can then store iron in
the form of ferritin and
hemosiderin
• Upon erythropoietin
stimulation of erythrocyte
synthesis, iron is released from
storage
• Iron binds to transferrin and is
carried to the bone marrow for
hemoglobin synthesis
Copper
• ☛ Important trace mineral that
Definition is a component of the plasma
proteins and ceruloplasmin
Fig. 3.1 -2. Oxidation of Ferrous Iron to Ferric
Iron via Hephaestin
Hephaestin • Transmembrane copper-
dependent ferroxidase
• Facilitates oxidation of Fe2+
(ferrous) to Fe3+ (ferric) for
proper transport of iron within
the intestinal lumen
• Enables iron to be absorbed
from the enterocytes to the
circulation in cooperation
with the basolateral iron
transporter ferroportin
• Major Cu++ carrier protein in
blood
Ceruloplasmin
• Facilitates transport of Cu++
• Has ferroxidase activity
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

Enables oxidation of iron from 3.2. ERYTHROPOIESIS

the ferrous to the ferric state
• Transport of iron for heme
synthesis decreases → iron
Copper accumulation in tissues
deficiency • Iron accumulating in the
tissues can lead to organ
damage
Zinc
• ☛ Trace mineral
• Coenzyme in the synthesis of
heme
• Coenzyme of the
aminolevulinic acid (ALA)
dehydratase that facilitates
the synthesis of the heme
portion of hemoglobin
• Upon erythropoietin stimulation, RBCs are
produced through a process called erythropoiesis
• ☛ RBCs develop from committed stem cells to
mature RBCs in about 7 days
• EPO will stimulate synthesis first of the
proerythroblast
o Organelles are still present
o Hemoglobin production begins
3.3. RED BLOOD CELLS (ERYTHROCYTES)
• RBCs have a remarkable ability to deform to
accommodate itself into small capillaries
• Due to the following factors:
o Biconcave disc-like configuration
o Membrane is strongly flexible and has
reversible deformation

Definition

Fig. 3.1 -3. Role of Zinc in ALA dehydratase

Folic Acid (Vit B9) and Cobalamin (Vit B12)
• Essential for DNA synthesis
Definition • Critical in the synthesis of new
cells, including the RBCs

Erythropoietin
• Released in response to tissue
hypoxia
• Cytokine glycoprotein
produced by the kidneys
Definition • Binding to membrane
receptors results in the
activation of JAK2 protein
kinases
• Commits myeloid progenitor Fig. 3.2 -1. Erythropoiesis
cells to differentiate into RBC (Instructor’s Slides)
• Regulated by the need to
deliver O2 to the peripheral
tissues
• Reduced tissue oxygenation
(e.g. anemia or hypoxemia) →
☛ Erythrocyte kidney releases the
Production glycoprotein-cytokine
erythropoietin → EPO binds
to and activates the JAK-STAT
pathway → stimulation of
proliferation and maturation of
erythroid progenitors

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Process of Erythropoiesis o ☛ Due to the absence of

• ☛ From the multipotent myeloid progenitor the organelles
cells, they commit to differentiate into • Requires erythropoietin
erythroid committed precursors stimulation for synthesis of
• The proerythroblast undergo morphologic new RBCs
changes:
o Cell size reduction
o Chromatin condensation
o Begins to produce hemoglobin
• Macrophages also start to interact with the
erythroblast to provide the iron to the cell
• Eventually the erythroblast will lose its
nucleus and some organelles forming two 3.3. RED BLOOD CELLS (ERYTHROCYTES)
daughter cells: reticulocytes and pyrenocytes
o ☛ Erythroblast undergo several cycles of
cell division where they lose some of
their organelles (e.g., golgi apparatus,
endoplasmic reticulum, and
mitochondria)
o Nucleus becomes smaller, more
condensed, and expelled out of the cell.
This process generates 2 daughter
structures: the reticulocytes and
pyrenocytes
▪ Reticulocytes - contains most of
the cytoplasm
▪ Pyrenocytes - contains the nucleus Functions
surrounded by a small cytoplasmic Fig. 3.3 -1. RBC Function
ring) (Instructor’s Slides)
▪ Most of the pyrenocytes are
• Deliver the maximum amount of
engulfed and degraded by the
oxygen that is needed by the
macrophages
tissues with the help of hemoglobin
• Retain their capacity to (present within the erythrocytes)
synthesize polypeptides (eg.
• Remove carbon dioxide, a waste
globin) because they have
product of cellular respiration, away
some ribosomes and
from the tissues and back to the
mRNAs
lungs
o ☛ Capable of limited • Contribute to acid-base
protein synthesis, such homeostasis
as synthesizing some
more of the cell's
Reticulocytes hemoglobin
• Spend 3 days in the bone
marrow and circulate 1-2
days before they become a
mature erythrocyte
o Thus, reticulocytes may
still be present in the
circulation comprising
around 1% of the total Structure
red cell population
• Devoid of organelles
o ☛ After 1 –2 days in the
blood circulation, most of
the reticulocytes Fig. 3.3 -2. RBC Structure
Mature eventually lose all of its (Instructor’s Slides)
Erythrocytes organelles become a
mature erythrocyte • Composed of a membrane
• Average life span in the surrounding a solution of
circulation: 100-120 days hemoglobin, which comprises 95%
• Unable to reproduce by itself

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of the intracellular protein of red
blood cells
• Have a remarkable ability to deform
in order to accommodate itself into
smaller capillaries. This due to one
of several factors:
1. Biconcave disc-like, round and flat
• High surface area-to-volume ratio
• Aids in rapid gas exchange within
the cell membrane
2. Membrane is strongly flexible and has
reversible deformation
• RBC diameter: 7 - 8 μm
• RBC cross-section
o Thickest: 2.0 – 2.5 μm
o Thinnest: 0.8 – 1.0 μm
• Capillary diameter: 3.5 μm
o Unique RBC cytoskeletal
network enables it to fold
over and squeeze through
the capillaries and
sinusoids of the spleen
3. Viscosity of the cytoplasm
• Dependent on intracellular
hemoglobin concentration
• Affects how the RBCs can
properly flow within the circulation
3.4. RBC MEMBRANE
Fig. 3.4 -1. RBC Membrane
(Instructor’s Slides)
• Composed of:
o Cytoskeleton
o Lipid Bilayer
• Includes:
o Phospholipids
o Sphingolipids
Lipid Bilayer
o Cholesterol
o Integral membrane proteins –
Band 3, Glycophorins
BIOCHEMISTRY
• Consists of spectrin tetramers
o Connected to the actin via
tropomyosin-tropomodulin
junctional complexes
• Tethered to the lipid bilayer via
o Immobile Band 3 proteins
o Adducin
At the spectrin-ankyrin
Band 3
binding sites
Forms bridge between
Cytoskeleton Adducin RBC bilayer and
cytoskeleton
• Plays an important role in RBC
membrane integrity
• Maintains biconcave, spherical
shape and flexibility in the
circulation
o Maximizes efficiency to
exchange O2 and C02
between RBCs and tissues
3.5. AGED RED BLOOD CELLS
Fig. 3.5 -1. Reasons for Aging
(Instructor’s Slides)
• Reasons for RBC aging:
• Oxidative damage from
o Cycle of gas exchange
Accumulate o Exposure to extrinsic
Oxidative chemicals
Damage • Oxidative damage accumulates in
the proteins within the RBC
o Induces cell aging
• Due to
o Cycles of osmotic swelling
and shrinkage
o Gradual loss of surface area
Cytoskeletal
→ decreases deformability
Damage
o Repetitive deformation
passing through small vessels
o Shearing forces from passing
through the heart valves
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• Absence of organelles → unable to o Broken down and removed

repair cytoskeletal proteins
o Also unable to synthesize
new hemoglobin
3.5.1. Fate of Aged RBCs
Heme
Fig. 3.5 -1. Removal of Aged RBCs
(Instructor’s Slides)
• Aged RBCs eventually die
• Removed through the spleen and liver
• RBC Lifespan: 120 days in circulation
o Removed from circulation by macrophages
• Macrophages
o Myeloid, phagocytic cells
o Located primarily in the bone marrow, liver,
spleen
• Hemoglobin components are further broken down,
recycled, or metabolized. Mainly into:
o Globin
o Iron
o Heme
from circulation via kidneys
• Stored in the liver in the liver or
spleen as
o Ferritin
Iron o Hemosiderin
• Carried by the blood stream by
transferrin to the bone marrow
o For recycling into new RBCs
• Non-iron portion of heme is
degraded into
o Biliverdin – green pigment
o Bilirubin – yellow pigment
• Bilirubin binds to albumin
o Travels via bloodstream to
the liver → utilized for bile
production
• In the large intestines
o Bacteria break bilirubin apart
from bile
o Converted to urobilinogen
and stercobilin → eliminated
in stool
• In the kidneys
o Circulating bilirubin and other
related byproducts (urobilin)
→ eliminated in urine

Fig. 3.5 -3. Pathways of Hemoglobin Metabolism and

Elimination
(Instructor’s Slides)

Fig. 3.5 -2. Hemoglobin Degradation Components

(Instructor’s Slides)

• Protein portion of hemoglobin

• Broken down into amino acids
Globin • Can be sent back to bone marrow
for production of new RBCs
• Hemoglobin not phagocytosed

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BREAKDOWN PIGMENTS • Glucose is taken up by the RBC via the

Fig. 3.5 -4. Hemoglobin Breakdown Pigments
(Instructor’s Slides)
• Breakdown pigments from hemoglobin destruction
can be seen in a variety of conditions
• In sites of injury
Bruising • Biliverdin produces dramatic colors
associated with bruising
• Due to a failing liver
• Bilirubin cannot be removed
Jaundice
effectively in the circulation
• Body assumes a yellow-tinged color
• Stercobilins – produces the typical
Stool and brown color associated with feces
Urine • Urobilins – produces yellow color of
urine
GLUT-1 transporter
o a.k.a. solute carrier family 2, facilitated
glucose transporter member 1
(SLC2A1)
o high affinity glucose transporter in the
membrane
o via facilitated diffusion
o NOT affected by insulin
• UTILIZATION OF THE GLUCOSE
SUPPLY OF RBC:
o 90% of glucose → glycolysis (EMP)
o 10% of glucose → HMP shunt (PPP)
4.1. EMBDEN-MEYERHOF PATHWAY (EMP) (RE0)
• Utilizes the majority (90-95%) of intracellular glucose
• Anaerobic process – glycolysis
• Generates ATP
o Preparatory phase – 2 ATP used
o Payoff phase – 4 ATP produced
o Net gain: 2 ATP per 1 mole of glucose
• Recall that RBC has no mitochondria – TCA cycle
(Krebs) & oxidative phosphorylation does not occur

4. RBC METABOLISM (5ML, RE0, SU1)

• Refers to the various metabolic pathways in the RBC

Lack organelles:
• No mitochondria – no TCA cycle,
electron transport chain, β-oxidation
pathway
• No nucleus – no nucleic acid synthesis
• No ribosomes – no protein synthesis
Require energy to maintain several
functions:
• ★ Maintenance of electrolyte gradient
between plasma and RBC cytoplasm
Mature through regulation of membrane ion
RBCs pumps (e.g., Na-K ATP, anion exchange)
• Synthesis of glutathione for its
protection against oxidative damage
o Protect Hgb, enzymes, cell membrane
• Maintenance of hemoglobin’s iron in its
ferrous (Fe2+) state
o Fe2+ state – functional reduced state
• Maintain the biconcave shape of RBCs
o Allow flexibility during blood circulation
and efficiency for gas exchange
Highly dependent on glucose for energy:

Fig. 4.1-1. Embden-Mayerhof Pathway

(Instructor’s slide)

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4.1.1. Preparatory Phase 4.1.3. Important Enzymes in EMP

• Otherwise known as the energy investment phase •
Exhibits the highest glycolytic
• Two ATP consuming steps: enzyme activity to isomerize
o Hexokinase DHAP to G3PO4
o Phosphofructokinase • G3PO4 can then enter the payoff
Triosephosphate
phase, undergoing the two ATP
Isomerase
generating steps
o This is why the ATPs
produced by the payoff
phase is 4 (see section 4.1)
• Transforms G3P to 1,3-BPG
with NAD as a cofactor
G3PO4
• Fixes Pi into high-energy
dehydrogenase
compound 1,3-BPG
• Reduces NAD to NADH

Fig. 4.1.2-1. Preparatory
(Instructor’s slide)
4.1.2. Payoff Phase
• Otherwise known as the energy generation phase
• Two ATP generating steps:
o Phosphoglycerate kinase
o Pyruvate kinase
• NADH generated in the G3PD step is utilized to
reduce pyruvate to lactate (see Fig. 4.1-1)
• This reduction of pyruvate to lactate through LDH
subsequently regenerates NAD that can be
reutilized in the G3PD step
PRODUCTS
• Regulator (regulatory intermediate) of
oxygen affinity to hemoglobin
1,3-BPG
• ☛ The two reactions [mentioned] favors
the synthesis of 1,3-BPG […]
• ★ Powers the cation exchange pump
ATP • ☛ [The two reactions] fix the metabolic
pathway to the generation of ATP
needed by the erythrocytes […]
• ★ Converts iron to its functional ferrous
NADH state
• ☛ [Reaction] reduces the NAD to NADH
✂ LECTURETTE/KAHOOT (LEAPNotes ‘25)
Why do red blood cells need to produce ATP?
A. For generation of oxidized glutathione
B. For conversion of ferric to ferrous
C. For Na-K ion pump
D. For synthesis of 2,3 BPG

Answer: C
RBCs want to generate reduced glutathione. Ferric to
ferrous conversion is facilitated by NADH or NADPH.
Synthesis of 2,3 BPG doesn’t really need ATP.

✂ LECTURETTE/KAHOOT (LEAPNotes ‘25)

Which metabolic pathway is present in the RBCs?
A. Embden-Meyerhof
B. Citric acid cycle
C. Oxidative phosphorylation
D. Beta Oxidation

Answer: A
We established that the mature RBCs do not have
mitochondria. As such, Krebs, ox phosph, & B-ox do not
happen in the RBC.

4.2. RAPOPORT-LUEBERING PATHWAY (RE0)

Fig. 4.1.2-1. Payoff
(Instructor’s slide)

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Fig. 4.2-1. Rapoport-Luebering Pathway
(Instructor’s slide)
• Unique branch of the glycolytic pathways seen in
mature RBCs
• Generates and dephosphorylates 2,3-
bisphosphoglycerate (2,3-BPG)
• Catalyzed by a single multifunctional enzyme
Bisphosphoglycerate mutase complex (BPGM) or 2,3-
BPG synthase/2-phosphatase

BPGM activity
• Isomerize 1,3-BPG to 2,3-BPG by
Mutase
bisphosphoglycerate mutase
• Hydrolysis of 2,3-BPG to 3-PG by
bisphosphoglycerate phosphatase
• ☛ Irreversibly dephosphorylates 2,3-
Phosphatase
BPG at C2 position
• ☛ 3-PG can reenter the main
glycolytic pathway

2,3-BPG
• Facilitates the supply of oxygen to the tissues by
binding to hemoglobin
• During periods when hemoglobin is deoxygenated,
the equilibrium is driven to the R-state → majority of
O2 will attach to Hgb (oxygen loading)
• In the presence of 2,3-BPG: it binds to the center
pocket of the 2 beta globin chain of Hgb → stabilizes
hemoglobin in the T-state → decrease O2 affinity to
Hgb (favor oxygen unloading).

Fig. 4.2-2. Effect of 2,3-BPG on Hgb

(Instructor’s Slides)

(Space intentionally left blank)

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SHIFT 3
Blood Generalities 17 JAN 2023 01
Dr. Judelyn T. Uy AY 2022-2023

ENERGETICS
• Metabolic flux through the Rapoport-Luebering shunt carries an energetic cost for the cell because it bypasses the
ATP-generating phosphoglycerate reaction (1st ATP-generating step of glycolysis)

Fig. 4.2-2. Bypassing of the Phosphoglycerate Reaction

(Instructor’s Slides)

• Acid pH or low ATP concentration in the RBC inhibits the mutase activity and activates the BPG phosphatase
and phosphoglycerate kinase activity

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Fig. 4.2-3. Effect of Acidity and Low Energy State on the Rapoport-Luebering Pathway
(Instructor’s Slides)

✂ LECTURETTE KAHOOT (RBC Metabolism) • Rapoport-Leubering is the shunt wherein

T/F: The Rapoport-Leubering shunt in the RBC is Bisphosphoglycerate Mutase (BPGM) generates
used to generate NADH? 2,3-BPG
• Importance of 2,3 BPG is for oxygen affinity, as it
Answer: FALSE favors oxygen unloading from Hgb molecules

Rationale:

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4.3. HEXOSE MONOPHOSPHATE SHUNT Hydrogen Donor NADPH

• Metabolizes 5-10% of total glucose influx to the RBC
• Important for the generation of NADPH (reducing
agent)
• Functions as cofactor in the:
o Reduction of oxidized glutathione to its
reduced sulfhydryl form: GSSG → GSH
o Reduction of iron to its ferrous state (to a lesser
extent): Fe3+ → Fe2+
Coenzyme FAD
• GSH (sulfhydryl form of glutathione): protects the RBC
from oxidative attack via the detoxification of toxic
peroxides (H2O2) and other toxic radicals (ROS)
within the RBC

Fig. 4.3.1-1. Glutathione and NADPH

(Instructor’s Slides)

4.3.2. Role of Reduced Glutathione

• Peroxides and ROS are normally eliminated by the
selenium-containing antioxidant enzyme: GSH
peroxidase
o Cofactor: Selenium
o Reactions:
o Reduces H2O2 → H2O
o Reduces Lipid Peroxides → lipid alcohols
Fig. 4.3-1. Hexose Monophosphate Shunt
Enzyme GSH Peroxidase
Cofactor Selenium
Reduction of:
H2O2 → H2O
Reactions Lipid Peroxides → Lipid Alcohols
Catalyzed
*Reduced glutathione is oxidized
back to its disulfide form
Sudden oxidant challenge diverts
glucose flow to the HMP shunt by
↑ ROS
20-30-fold that its usual metabolic
activity
Cells with lower levels of reduced
glutathione are more susceptible
↓ GSH
to oxidative attack → membrane
damage & hemolysis
Fig. 4.3-2. NADPH Production
(Instructor’s Slides) 4.3.3. Effect of NADPH to Hemoglobin
• Iron is vital to Hgb because it facilitates the binding of
O2 to molecule.
4.3.1. Effect of NADPH to Glutathione • Iron should be in its reduced ferrous form (Fe2+) to
• GSSG can be reduced through disulfide cleavage → reversibly bind to the heme groups of the Hgb molecule.
Sulfhydryl form

Enyzme GSH Reductase

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Fig. 4.3.3-1. Ferrous Iron in Hemoglobin

(Instructor’s Slides)

• When the iron atom in the heme group of Hgb becomes

oxidized (lose an electron): Ferrous (Fe2+) → Ferric
(Fe3+)
o Hemoglobin → Methemoglobin
(Ferrihemoglobin)
o Typical bright red blood → Chocolate brown
• Methemoglobin: one or more heme irons were
oxidized to its ferric state
o Does NOT bind oxygen
o Oxidized ferric iron is susceptible to generate
ROS by reducing oxygen to superoxide
radical
o Thus, it is important to reduce the iron back to
its reduced ferrous state
Fig. 4.3.3-2. Methemoglobin
(Instructor’s Slides)
Fig. 4.4-1. Methemoglobin Reduction Pathways
Major Pathway: Blue highlight below
Minor Pathway: Violet highlight above
(Instructor’s Slides)
4.5. CARBONIC ANHYDRASE
• RBC has high levels of carbonic anhydrase
• Carbonic anhydrase plays an important role in
respiration by influencing carbon dioxide transport in
the blood
• CO2 has low solubility in aqueous solution
• Once CO2 (from the tissue) is absorbed by the RBC,
carbonic anhydrase catalyzes conversion of CO2 to
carbonic acid (H2CO3) and bicarbonate (HCO3-)
o Increases solubility of CO2 resulting in rapid
absorption from cells and expulsion in the lungs
• Once the RBC reaches the alveoli, this same enzyme
will help convert the bicarbonate ions back to CO2
for proper expiration in the lungs
o Without this enzyme, very little CO2 will be
transported in the blood away from the tissues
o

4.4. METHEMOGLOBIN REDUCTION PATHWAY

• In order to regenerate hemoglobin and maintain a low
level of methemoglobin in the blood, two mechanisms
are proposed:

NADH-Cytochrome B5
Major Methemoglobin Reductase
Pathway • NADH source: Glyceraldehyde-3-
Phosphate Dehydrogenase Reaction
NADPH Methemoglobin Reductase
Minor • Requires an exogenous electron
Pathway acceptor (e.g., methylene blue)
• NADPH source: HMP Shunt
Fig. 4.5-1. Carbonic Anhydrase
(Instructor’s Slides)

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✂ LECTURETTE KAHOOT (RBC Metabolism) • Both enzymes function to reduce iron from ferric to
Which metabolic pathway is not available in the RBC ferrous; ferrous iron is good for hemoglobin as it
as a source of ATP production? (1 or more answers favors binding more to oxygen as compared to its
possible) ferric hemoglobin/ferric methemoglobin
A. Beta-oxidation • Embden-Meyerhof (glycolysis): ATP proudction
B. Kreb’s cycle • Rapoport-Luebering: 2,3-BPG generation
C. Anaerobic glycolysis • Pentose Phosphate: NADPH production
D. Hexose monophosphate shunt • Additional: Hephaestin can convert ferrous iron to
ferric, usually iron is transported with ferroportin
Answer: A, B, D protein from the intestinal membrane; the ferric iron
is brought to the circulation through transferrin (wants
Rationale: iron to be in ferrous form)
• A & B - the RBC has no organelles, including the • In the circulation iron is in ferric form but once in Hgb,
mitochondria (in which beta oxidation & Kreb’s cycle it must be in ferrous form
take place) What is the importance of carbonic anhydrase
• D – HMP shunt is for NADPH production (not ATP) present in the RBC?
Glucose enters the RBC thru: A. Keep iron in its ferrous state
A. GLUT 4 B. Converts CO2 to more soluble HCO3
B. GLUT 3 C. Needed by NADH cytochrome BS reductase
C. GLUT 1 D. Generate NADPH used to reduce glutathione
D. GLUT 2
Answer: B
Answer: C
Rationale:
Additional Note: GLUT1 is NOT insulin dependent • RBCs have a high amount of carbonic anhydrase
Which is TRUE of metabolism occurring with the because it tries to break CO2 into carbonic acid [so
RBC? that] it will be more soluble for the RBC to carry before
A. Generation of ATP occurs through TCA cycle and ETC it can bring it back to the lungs where this enzyme will
B. Bisphosphoglycerate mutase generate ATP needed convert it again to CO2
by Na-K pump
C. NADP dependent oxidation of GSH protects against 4.6. BIOCHEMICAL BASIS OF ABO BLOOD GROUP
oxidative attack SYSTEM
D. LDH generates NAD+ needed by the glyceraldehyde
PO4 dehydrogenase

Answer: D

Rationale:
• A – RBC has no mitochondria
• B – Bisphosphoglycerate generates 2,3-BPG
• C – should be REDUCTION
• D – Pyruvate → Lactate is the last reaction of
anaerobic glycolysis; important for RBC because it
will help generate the NAD+ to be utilized again by
the Step 6 of glycolysis (conversion of
glyceraldehyde-3-phosphate to 1,3-BPG)
Which metabolic pathway facilitates reduction of Fig 4.6-1. Different Blood Group Antigens in RBC Membrane
ferric to ferrous allowing better O2 carrying capacity (Instructor’s Slides)
of red
cells? • ☛ The ABO blood group antigens are characterized by
A. Embden-Meyerhof different polysaccharide chains attached mainly on a
B. Methemoglobin reductase glycolipid on an RBC membrane.
C. Rapoport-Luebering o Can be classified as type A, B, O, AB
D. Pentose phosphate

Answer: B

Rationale:
• Methemoglobin reductase has 2 enzymes: NADH
cytochrome b5 reductase and NADPH
methemoglobin reductase
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o Leads to the formation of 3 distinct substances:

▪ H substance
▪ A substance
▪ B substance

Fig. 4.6-2. ABO Blood Group System

A, B, AB, O (from top left to bottom right)
(Instructor’s Slides) Fig 4.6-5. Initial Formation of Four Sugar Moieties
(Instructor’s Slides)

• Membranes of RBCs of each individual contain one

blood group substance. • Initial formation of the first 4 sugar moieties that attach
to ceramide is formed by the sequential action of distinct
glycosyltransferases
o Incorporates in this particular order:
1. Glucose
2. Galactose
3. N-acetylglucosamine
4. Galactose

4.6.1. H Substance

Fig 4.6-3. Common ABO Antigen Structure

(Instructor’s Slides)

• ABO antigens are complex oligosaccharides.

• In RBC membranes, this ABO substances are present
in glycosphingolipids.
o ☛ Particularly joined by a glycosidic bond
between glucose and the hydroxyl group in
carbon 1 of sphingosine.

Fig 4.6.1-1. Formation of H Substance

(Instructor’s Slides)

• Formed by alpha 1,2

fucosyltransferase attaches
fucose to the terminal galactose
residue.
Formation o ★ ABO blood group
antigen synthesis begins
with this enzyme
Fig 4.6-4. Different Oligosaccharide Groups in ABO Antigens o Precursor of both A and B
(Instructor’s Slides) substance
Antigen NO A and B antigen
• The blood types are differentiated by the
Blood Type Type O – “universal donor”
oligosaccharide groups attached to the sphingolipid
(ceramide)
4.6.2. A Substance
• There are 5 sugar residues responsible for forming
specific chains – glucose, galactose, fucose, N-
acetylglucosamine, and N-acetylgalactosamine.
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES
Fig 4.6.2-1 Formation of A Substance
• Formed by alpha 1,3
acetylgalactosamine (GalNac)
transferase.
Formation o Attaches
acetylgalactosamine to
the terminal galactose of
the H substance
A antigen/Anti-A antibodies are
Antigen
directed against GalNac residue
Blood Type Type A – NO anti-A antibodies
4.6.3. B Substance
Fig 4.6.3-1 Formation of B Substance
• Encodes UDP-Gal specific
galactosyltransferase (alpha 1,3
Formation galactosyltransferase)
o Adds another galactose
residue to the H substance
B antigen/Anti-B antibodies are
Antigen
directed against Gal residue
Blood Type Type B – NO anti-B antibodies
4.6.4. A and B Substance
BIOCHEMISTRY
N-
Fig 4.6.4-1 A and B Substance
N-
• Biantennary structure → contains
one type A chain and one type B
chain (2 arms of the
oligosaccharide)
• Capable of both A and B substance
Formation
in RBC membrane
o One has terminal N-
acetylgalacosamine and
the other having the
terminal galactose residue
A and B antigen WITHOUT Anti-A and
Antigen
Anti-B antibodies
Blood Type Type AB – “universal recipient”
✂ ABO BLOOD GROUP (Kahoot)
What is the ABO blood type of the sugar residue that
contains 1-Glu, 2-Gal, 1-GluNac,1-Fucose?
Answer: Type O (H Substance)
• H substance is Type O blood
• Type O blood contains 1-Glu, 2-Gal, 1-GluNac, and
1-Fucose
• H substance is different from A and B substance in a
way that there’s only 5 carbohydrates. However, the
figure above only shows 4. Dr. Uy said that the image
is lacking a carbohydrate residue.
• A and B substance have an additional carbohydrate
residue
• B substance has 3 galactose units with the additional
Galactose included
• A substance has GalNac as the added carbohydrate
In which molecule of the RBC membrane are the ABO
blood group substances seen?
• Sphingolipids
• Glycophorins
• Spectrin
• Ankyrin
Answer: Sphingolipids
• Sphingolipids are complex lipids that are
components of the RBC membrane wherein the ABO
blood group substances (carbohydrate
21
SHIFT 3| LESSON 1 | BLOOD GENERALITIES

oligosaccharides) are attached to in order to identify Abnormality in

• Hereditary spherocytosis
the blood group. cytoskeletal
• Hereditary elliptocytosis
• Ceramide (Terminal Carbon 1) would be able to bind proteins
oligosaccharides – usually found in red cell
membrane wherein it is exposed to the surface and 6. PLATELETS (THROMBOCYTES) (8LJ)
indicate whether you are blood type O, A, B, or AB ● Comprise <1% of the cellular elements of blood
Which enzyme is responsible for the formation of B
antigen?
• Galactosyltransferase
• Fucosyltransferase
• Carbonic anhydrase
• GalNac transferase

Answer: Galactosyltransferase
• Galactosyltransferase – galactose is the added
carbohydrate in B antigen
• GalNac transferase – GalNac is the added
carbohydrate in A antigen
• Fucosyltransferase – adds fucose to the H antigen

5. DISORDERS AFFECTING RBC (SU1)

DISORDER MAJOR CAUSE

Iron deficiency Inadequate intake or excessive
anemia loss of iron
Megaloblastic Deficiency in Vit. B12 or Folic
anemia acid Fig 6-1 Formed elements of blood
Intake of excess oxidants (Instructor’s slide)
(sulfonamides or aniline)
● Synthesized from common myeloid precursor
Genetic deficiency of NADH ○ ☛ Stimulation by thrombopoietin → production &
Methemoglobinemia cytochrome b5 reductase differentiation of megakaryocytes in the bone
marrow → fragment: 1000-4000 platelets per
Inheritance of hemoglobin M megakaryocyte
(mutation in histidine residues of ● Thrombopoietin - cytokine that stimulates the bone
Hgb) marrow megakaryocytes to fragment and form platelets
Genetic condition that causes ○ A glycoprotein produced primarily by the liver
Hereditary the body to absorb excessive
hemochromatosis quantity of iron (tissue damage
due to superoxide
Mutation in the gene for G6PD
(HMP shunt
G6PD deficiency
Inability to produce NADPH and
reduce GSH increased oxidative
stress → hemolytic anemia
Pyruvate kinase RBC isozyme PK gene mutation
deficiency decreased ATP production
Hereditary
Abnormalities in the amount and
spherocytosis/
structure of cytoskeletal proteins
elliptocytosis

CAUSE DISORDER Fig 6-2 Thrombopoietin

• Iron deficiency anemia
Exogenous causes • Megaloblastic anemia
• Methemoglobinemia
• Methemoglobinemia
Aberrations of • Hereditary hemochromatosis
metabolic activities • G6PD deficiency
• Pyruvate kinase deficiency
BIOCHEMISTRY
(Instructor’s slide)
● Small, irregularly shaped, colorless fragments
● Size: 1-4 μm, about 20% of the diameter of RBCs
● Average life span: 8-9 days
● Normal platelet count: 1150,000-350,000/uL
● ☛ Make up only 1% of fraction of formed elements
because they are very small
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

6.1.1. 3 Types of Granules

GRANULE TYPES AND CONTENT

• Calcium
(Electron) • ADP
Dense • ATP
• Serotonin
• Heparin Antagonists
• Platelet Derived Growth Factor,
Fig 6-3 Thrombocytes
• Thromboglobulin
Alpha
(Instructor’s slide) • Fibrinogen
• von Willebrand Factor
6.1. FUNCTION OF PLATELETS (THROMBOCYTES) (8LJ) • Other clotting factors
● Maintain homeostasis – principal function is to prevent Lysosomal • Hydrolytic Enzymes
bleeding; depends on 3 factors
○ Endothelial supporting function
○ Formation of platelet plug (mechanical plug at site
of vessel injury)
○ Secretion of coagulation factors (regulators of
clotting factors and vascular repair)

Fig 6.1-1 Function of thrombocytes

(Instructor’s slide)

● Not involved in any vascular injury = inactive

○ Usually invaginates into the interior of the cell = Fig 6.1-3 3 Types of granule and what they contain
open membrane canalicular system (Instructor’s slide)
○ Canalicular structure substantially increases the
membrane surface area that is potentially available 6.2. MAJOR METABOLIC PATHWAYS (8LJ)
for clotting reactions and facilitate secretion of ● Main energy source: Glucose
various endocrine and coagulation factors ● Energy source for aggregation: glycolysis,
● Also contain an extensive actin-myosin system glycogenolysis
○ Upon platelet activation in response to endothelial ● Mitochondria
injury → calcium-dependent changes in the ○ Allow platelets to Generate ATP from TCA cycle
contractile elements = long pseudopods on the and ETC and B-oxidation of fatty acids (secondary
membranes of the platelets favoring their adhesion source of energy)
to injured blood vessels

Fig 6.1-2 Active and inactive platelets

(Instructor’s slide)

● NO nucleus thus do NOT reproduce Fig 6.2-1 Content of a thrombocyte

○ ☛ Unlike RBCs they possess mitochondria and (Instructor’s slide)
three types of granules
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6.3. DISORDERS AFFECTING PLATELETS (8LJ) ● The 5 major types of leukocytes are further divided into
2 main groups
PLATELET DISORDERS
Genetic mutation that
impairs the ability of
Von Willebrand
platelets to adhere to the
Disease
endothelium and deficiency
of Factor 8
Bernard-Soulier Inherited deficiency in
Syndrome glycoprotein Ib
Inherited deficiency in the
Glanzmann
glycoprotein IIb/IIIa
Thrombasthenia
complex
Characterized by formation of
Acute Coronary enlarged, hyperactive
Syndrome
platelets → thrombosis
Immune Depressed platelet count Fig 7-1 Granulocytes and agranulocytes
Thrombocytopenic due to autoantibodies on (Instructor’s slide)
Purpura platelets
Mutation on the glycoprotein 7.1. MAIN TYPES OF LEUKOCYTES (8lJ)

IIb/IIIa generates
Alloimmune
Thrombocytopenia
alloantibodies that attacks 7.1.1. Granulocytes
both endogenous and
● Neutrophils, eosinophils, basophils
donated platelets
Progressive kidney failure ● ☛ Possess granules in their cytoplasm
Hemolytic-Uremic
results to thrombocytopenia ● ☛ Nuclei are often segmented or multinucleated
Syndrome ● ☛ Granules contain many cell-signaling molecules
and hemolytic anemia
that mediate inflammatory processes
7. WHITE BLOOD CELLS (LEUKOCYTES) (8LJ)
● Part of the immune system 7.1.2. Agranulocytes
o Potent defenders against invading pathogens ● Monocytes, lymphocytes
o Participates in the acute inflammatory response ● ☛ Do NOT possess granules in their cytoplasm
● Comprise <1% of the cellular blood
● ☛ Usually have a large irregularly shaped nucleus
● WBC count: 4,000-10,000/µL
7.2. ORIGIN OF LEUKOCYTES (HAE)

☛ COMMON MYELOID PROGENITOR

Fig 7-1 Major white blood cell types

(Instructor’s slide)

● ☛ Generally larger than RBCs Fig. 7.1-1. Stimulation of GM-SCF, producing monocytes and
● Possess complete organelles granulocytes
● Nucleated cells (Instructor’s Slides)
o ☛ Nuclei are unique because of the marked • Some WBCs come from the common myeloid
deviation from the usual nucleus of most eukaryotic precursors in the bone marrow
cells

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• Upon stimulation by the granulocyte monocyte- • Basophils and Neutrophils – Relatively short life
colony stimulating factor (GM-SCF), spans
differentiation and maturation can produce: • Lymphocytes – May live for years as memory cells
o Monocytes
o Granulocytes 7.4. NEUTROPHILS (HAE)
▪ Neutrophils,
▪ Eosinophils NEUTROPHILS
▪ Basophils

☛ COMMON LYMPHOID PROGENITOR

Fig. 7.4-1. Neutrophils

(Instructor’s slide)

• Also known as Polymorphonuclear cells (PMNs)

because of their multisegmented nucleus.
• Most abundant type of leukocytes

FUNCTION OF NEUTROPHILS
• Usually the first responders to microbial infection
Fig. 7.1-2. Stimulation of GM-SCF, producing monocytes and • Mainly defend against bacterial and fungal
granulocytes infection
(Instructor’s Slides) o Initiates respiratory burst through
• Lymphocytes come from the common lymphoid phagocytosis
progenitor o Release of toxins
• Upon stimulation by specific cytokines → give rise to ▪ Hydrolytic enzymes
small lymphocytes ▪ Reactive oxygen species
o B cells → mature in the bone marrow ▪ Antimicrobrial peptides
o T lymphocytes → migrate and mature in • Encapsulate invading bacteria and fungi within
the thymus membrane vesicles through phagocytosis
• Their activity and death in large numbers from
7.3. GENERAL CHARACTERISTICS OF LEUKOCYTES (HAE) degranulation form purulent necrosis (pus)
NEUTROPHILS EOSINOPHILS BASOPHILS MONOCYTE LYMPHOCYTE
7.5. EOSINOPHILS (HAE)

EOSINOPHILS

60-70% 2-4% 0-1% 5% 25%

12-15 μm 12-15 μm 12-15 μm 12-20 μm 6-18 μm
Few
May live
7 hours 8-12 days hours to 3 days
for years
few days
(A larger copy of this table is attached in the appendix)

• Neutrophils – Most abundant Fig. 7.5-1. Eosinophils

• Basophils – Least abundant (Instructor’s slide)
• Monocytes – Largest in size
• Lymphocytes – Smallest in size FUNCTION OF EOSINOPHILS
• Primarily deal with parasitic and viral infections
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

• Phagocytize parasites • Consume and destroy invading organisms

• Remove fibrin during inflammation
• Participate in immediate allergic reactions
• Modulate allergic inflammatory responses
7.6. BASOPHILS (HAE)
• Consume necrotic host cells that have been
compromised by infection or have been left behind
by granulocyte attack
• Remove damaged/aged RBCs
o Helps maintain oxygen-delivering capability
of erythrocytes

BASOPHILS 7.8. LYMPHOCYTES (HAE)

LYMPHOCYTES

Fig. 7.6-1. Basophils

(Instructor’s slide)

• Release heparin, histamine, and chemokines

Fig. 7.8-1. Lymphocytes
• Involved in allergic/hypersensitivity (Instructor’s slide)
reactions
• Main cells of the immune system and are the primary
• Produced from the decarboxylation of
antigen-recognizing cells
Histidine
Histamine
• Stored in secretory granules
TYPES OF LYMPHOCYTES
• Can cause vasodilation and promote
• Make up 75% of the lymphocytes
blood flow to the infected tissues,
causing an allergic reaction
T Cells • Participate in cell-mediated
immunity
Heparin • Blood thinning substance
• Make up 10% of the lymphocytes
FUNCTION OF BASOPHILS • Generate and release protective
B Cells antibodies that targets foreign
• Defend against parasites and bacteria by releasing
invaders and tag them for elimination
histamine
• Participate in humoral immunity
• Releases chemokines that attract additional
• Cytotoxic cells that participate in the
neutrophils to sites of infection
Natural innate immune response against
Killer (NK) viruses and tumor cells
7.7. MONOCYTES (HAE)
Cells • Mediates fever and long-lasting
inflammation
MONOCYTES

7.9. MAJOR METABOLIC PATHWAYS (HAE)

Fig. 7.7-1. Monocytes and Macrophage Fig. 7.9-1. Leukocyte Organelles

(Instructor’s slide) (Instructor’s slide)

• Largest type of Leukocyte • Major metabolic pathways

• Migrate from the blood stream to diseased tissues o Aerobic Glycolysis
• Under varying conditions, they differentiate into o Oxidative phosphorylation
phagocytic macrophages, dendritic cells, or foam o Pentose Phosphate Pathway
cells to engulf infected and damaged host cells • Note that Leukocytes contain all organelles that are
present in other cells.
MACROPHAGES
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GENERALITIES 7.10.2. Diapedesis

FUNCTION PATHWAY INVOLVED • Once the leukocytes have migrated to the site of injury,
Energy • Aerobic Glycolysis adhesion to the endothelial cell is mediated by
Generation • Oxidative Phosphorylation transmembrane glycoproteins integrin and selectin
• Pentose Phosphate Pathway families
Antioxidant
and NADPH
o NADPH are needed by • Migration of the leukocytes out of the circulation via
leukocytes during the amoeboid mechanism
Production
respiratory burst o Gradually squeezes out the contents of the cell
through pseudopod projection in between the
7.10. LEUKOCYTE ACTIONS (HAE) cells of the capillary endothelium

7.10.1. Chemotaxis 7.10.3. Phagocytosis

Fig. 7.10.1-1. Cleavage of PIP2 and IP3-DAG
(Instructor’s slide)
• Chemotaxis is the migration of leukocytes to sites of
injury in response to chemical signals (chemotactic
factors) that bind to G-protein coupled receptors
o Chemokines – stabilized by disulfide bonds
o Complement fragment C5a
o Small peptides & Leukotrienes
• Chemotactic factors can induce a signal-transduction
cascade that will activate phospholipase C
• Activation of the phospholipase C will hydrolyze the
phosphoester bond between the phosphate group and
OH group of the alpha prime carbon of the glycerol
backbone of phosphatidylinositol-4,5-bisphosphate
(PIP2)
• This will yield diacylglycerol (DAG) and the second
messenger, inositol-1,4,5-triphosphate (IP3)
o IP3 → mobilizes the calcium stores to activate
the actin and myosin cytoskeleton of the
neutrophils and promote migration and granule
secretion
• Leukocytes destroy invading microorganisms through
phagocytosis
• Mediated by phagocytic neutrophils, eosinophils,
and macrophages
• These cells often recognize and bind to foreign invading
microorganisms either by using:
1. Receptors that recognizes bacterial LPS or
peptidoglycans
2. Presence of antibodies or complement
factors
• Once the phagocytic cells binds to invading organisms,
they are internalized into membrane vesicle called
phagosome.
• This phagosome will then bind with the granules
present within the cells that will secrete hydrolytic
enzymes like lysozymes, proteases, anti-microbial
peptide (defensins), and reactive oxygen species that
can destroy and digest the microbial invader.
• The debris left after digestion will eventually be expelled
out of the cell.
Fig. 7.10.3-1. Phagocytosis
(Instructor’s slide)

7.10.3.1 Phagocytic Granules

• Phagocytic leukocyte posses in their granules several enzymes or proteins that facilitates digestion of foreign
microorganism.
• Granule-derived enzymes and cytotoxins fuse with the phagolysosome to destroy foreign pathogens

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Reaction Catalyzed or
Enzyme or Protein Notes
Function
H2O2 + X- (halide) + H+ → HOX • ☛ Catalyzes the synthesis of hypochlorous or other
+ H2O hypohalous acids that are powerful oxidants and
Myeloperoxidase (MPO) microbicidal to invading organisms
where X = Cl-, HOX = • Responsible for the green color of pus
hypochlorous acid • Genetic deficiency can cause recurrent infections
2O2 + NADPH → 2O2-. + NADP • ☛ Also microbicidal to invading microorganisms by
+ H+ generating superoxide that eventually can also form
hydrogen peroxide
*O2-. – oxygen radical • ☛ This enzyme system also increases the
consumption of both oxygen and NADPH by the
NADPH Oxidase WBC.
o NADPH from the HMP shunt
o High oxygen consumption results into
respiratory bursts
• Key component of the respiratory burst
• Deficient in chronic granulomatous disease
Hydrolyzes link between N- • Abundant in macrophages
Lysozyme
acetylmuramic acid and N- • Hydrolyzes bacterial peptidoglycans
acetyl-o-glucosamine found in
certain bacterial cell walls
Basic antibiotic peptides of 20- • Apparently kill bacteria by causing membrane
Defensins
33 amino acids damage
Iron-binding protein • May inhibit the growth of certain bacteria by binding
Lactoferrin* iron and may be involved in regulation of
proliferation of myeloid cells
Elastase Proteases • Abundant in phagocytes
Collagenase • Breakdown protein components of infectious
Gelatinase organisms
Cathepsin G • Generate fragments for antigen presentation
*Not emphasized

7.10.4. Opsonization
• Process of tagging pathogen with proteins to facilitate
recognition by phagocytic cells

7.10.5. Neutrophil extracellular traps (NETs)

• Neutrophils and Eosinophils can eliminate larger
pathogens by trapping them within webs called
Neutrophil extracellular traps (NETs).
• These nets are formed by the dispersal of the chromatin
strands (strands of chromosomal DNA) of the multi-
lobed nucleus, and with it, the membranes of the
Fig. 7.10.5-1. Neutrophil extracellular trap formation and
granules also rupture and allows binding of all its release
enzymes and proteins and cytotoxins to the chromatin (Instructor’s slide)
strand, generating a mesh-like net.
• Once cell lysis, it will then release the DNA protein web ✂ LECTURETTE KAHOOT (Granule-derived
to serve as traps for invading parasites. Enzymes)
Present in the granules of phagocytic cells and
destroys invading pathogens by releasing
hypohalous acids:
• Defensin
• Myeloperoxidase
• Proteinase
• NADPH Oxidase

Answer: Myeloperoxidase
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Rationale:
• Myeloperoxidase can generate hypohalous
acid, which is responsible for destroying
infections. It is also responsible for the green
color of pus.
• NADPH Oxidase induces production of reactive
oxygen species (ROS) thus resulting to a
respiratory burst of the organism.
• Defensins are capable of damaging the
membranes of invading pathogen
• Proteases breakdown protein components of
infectious agents to kill them

7.11. DISORDERS AFFECTING WHITE BLOOD CELLS (WUT)

DISORDER MAJOR CAUSE

Malignant neoplasms of blood-
Leukemias forming tissues or any WBC
components
Lack of 2 subunit of Integrin
LFA-1, Mac-1, and p150,95 →
impairs leukocytes to adhere to
Type 1 Leukocyte
vascular endothelial cells in the
Adhesion
Deficiency site of injury
Patients having this condition
suffer from recurrent infections
Mutations in the genes encoding
the NADPH oxidase system →
impaired production of ROS that
Chronic
can kill invading pathogens
granulomatous
disease
Patients having this condition
also suffer from recurrent
infections
8. PLASMA (C33)
● Slightly yellow straw-colored liquid
● Comprises of 55% of the total blood volume
● 91% of plasma volume is water
● 7% of plasma volume are proteins
● ☛ Remaining 2% contains a complex mixture of
electrolytes, nutrients, vitamins, waste product,
hormones, clotting factors and proteins
Fig. 8.1-1. Plasma
(Instructor’s slide)
9. PLASMA PROTEINS(C33)
● Functions:
○ Transport of nutrients, enzymes, hormones, and
proteins (main role)
○ Regulation of osmotic pressure
○ Maintain fluid and electrolyte balance
○ Involved in inflammatory response, immune
defense, blood clotting, & complement cascade
9.1. EFFECT OF PLASMA PROTEINS IN OSMOTIC
PRESSURE(C33)
Exerted by the amount of plasma
proteins that do NOT leave the capillary
Oncotic and draw water
Pressure
o ☛ Pulls water back into the
circulation
Based on the pressure exerted by the
blood pushing against the walls of the
Hydrostatic capillaries
Pressure
o ☛ Brings the fluid out into the
interstitial fluid compartment
● Since the osmotic pressure affected by the plasma
proteins are constant in both the arterioles and
venules, the effect exerted by hydrostatic pressure
differences in the arteries and veins affect how fluid
goes in and out of the capillaries

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Fig. 9.2-1. Sizes of Plasma Proteins

Fig. 9.1-1. Fluid Balance (Instructor’s slide)
(Instructor’s slide)
THREE MAJOR GROUPS
● Such that, higher hydrostatic pressure than the Albumin • Most abundant and smallest in
osmotic pressure in the arteries, promote fluid to flow (54-60%) terms of size & molecular mass;
out into the extravascular space • Alpha (α)-1 globulin – 3%
● While lower hydrostatic pressure than osmotic Globulin • Alpha (α)-2 globulin – 7%
pressure in the venules favors fluid to move back into (35-38%) • Beta (β) globulin – 12%
the capillaries • Gamma (γ) globulin – 16%
Fibrinogen
---
(4-7%)

Fig. 9.1-2. Fluid Balance Fig. 9.2-2. Proportion of Plasma Proteins

(Instructor’s slide) (Instructor’s slide)

● In conditions where the concentration of plasma 9.3. ALBUMIN(0NY)

proteins decreases, osmotic pressure will eventually
go down even below the hydrostatic pressure in the
venules resulting to fluid flowing out to the
extravascular spaces in the entire circulation and
clinically manifests as edema
9.2. PLASMA PROTEINS OVERVIEW(C33)
● Comprises the major part of the solids of plasma (7%)
● Concentration: 7.0 – 7.5 g/dL
● 70-80% synthesized by the liver EXCEPT for:
o vonWillebrand factor (vWF) by vascular
endothelium
o γ-globulins by lymphocytes/ plasma cells
● Rich in disulfide bonds
● Contain bound carbohydrate (glycoproteins) or lipid
(lipoproteins)
• Major plasma protein (60%)
o To make a mature albumin, there is a need to
cleave off a single peptide and hexapeptide in
the n-terminus during synthesis
• A single polypeptide chain with 585 amino acids
o Folded into 3 structural domains
o Stabilized by 17 intrachain disulfide bonds
• Low molecular weight (69 kDa)
• Responsible for 75-80% of the oncotic pressure
• At pH 7.4, characterized as an anion with 20 negative
charges per molecule
• Has a high capacity for nonselective binding for many
ligands
• Blood conc: 35-45 g/L
• Plasma half-life (T1/2) = 20 days

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Decrease albumin synthesis

Kwashiorkor due to protein malnutrition

✂ LECTURETTE/KAHOOT (LEAPNotes ‘25)

Which of the following is not a function of albumin?
A. Oncotic pressure
B. Clot formation & wound healing
C. Esterase activity
D. Transport drugs and hormone

Answer: Clot formation & wound healing

Which plasma protein contributes the most to the
osmotic pressure of the blood plasma?

A. Albumin
B. Globulin
C. Fibrinogen
D. Ceruloplasmin
Fig. 9.3-1. Albumin with many polar groups
(Instructor’s slide) Answer: Albumin
• Other globulin fractions that can contribute to
FUNCTIONS osmotic pressure: alpha and beta globulins
• Regulation of osmotic pressure – major function
• Bind and transport numerous ligands (contains 7 9.4. FIBRINOGEN(0NY)
fatty acid binding sites) • Large glycoproteins
• Free fatty acids, sterols • 4-7% of the plasma proteins
• Drugs: salicylates, barbiturate, sulfonamides, • Functions: clot formation, wound healing
• penicillin & warfarin
• Hormones: sex hormones, thyroid hormones
• Calcium, copper zinc
• Bilirubin
• Esterase activity

Fig. 9.3-2. Albumin binding sites Fig. 9.4-1. Different plasma proteins
(Instructor’s slide) (Instructor’s slide)

DISORDERS LOWERING ALBUMIN LEVELS IN BLOOD 9.5. GLOBULINS (0NY, KSH)

Genetic mutation that impairs • Heterogenous group of globular proteins
Analbunimeia the ability to synthesize • 2nd to albumin in volume and have a higher in
albumin molecular weight than albumin
Liver disease Decrease albumin-globulin • 35-38% of plasma proteins
(cirrhosis) ratio • Subgroups:
Kidney disease o Alpha globulins (ɑ1& ɑ2)
(nephrotic o Beta globulins (β)
syndrome) o Gamma globulins (γ)

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✂ LECTURETTE/KAHOOT (LEAPNotes ‘25)

Which globulin fraction/s can facilitate transport of
ions, hormones, & hemoglobin and contribute to the
osmotic pressure?

A. Alpha Globulins
B. Beta Globulins
C. Gamma Globulins

Answer: A and B
Gamma globulins or immunoglobulins serve to protect
our body from diseases (primary function). Alpha and
Beta globulins are produced by the liver while the gamma
globulins are produced by the plasma cells or
lymphocytes.

9.5.1. Immunoglobulins/Gamma globulins Fig. 9.5.1-2. Antigen-antibody complex

• Glycoproteins (Instructor’s slide)
• Aid in immune response
• Specifically recognize and bind to antigens (e.g., 9.5.2. Alpha globulins
bacteria, viruses) and aid in their destruction
• Various classes differ in structure, target specificity, TYPES OF ALPHA GLOBULINS
biological features, and distribution ALPHA1 GLOBULINS ALPHA2 GLOBULINS

α1-Lipoprotein
α1-Fetoprotein
α1-Antitrypsin (α1-
antiproteinase) Pre-β lipoprotein
α1-Acid glycoprotein Ceruloplasmin
(orosomucoid) Α2-Macroglobulin
Retinol-binding protein Haptoglobulin
Thyroxin-binding protein
Transcortin (corticosteroid-
binding protein)

Fig. 9.5.1-1. Immunoglobulins

(Instructor’s slide) ALPHA-1 GLOBULINS
• Major glycoprotein in fetal plasma
• Formation of antigen-antibody complex → mediates & amniotic fluid
phagocytosis, inflammation, cell lysis α1-fetoprotein • Levels increase during pregnancy
• Specific proteases degrade & remove these
• Tumor marker for HCCA or
complexes to prevent further inflammatory responses
teratoblastomas
that can damage normal tissues
• AKA High-Density Lipoprotein
α1-lipoprotein • Delivers cholesterol back to the
liver (“Good cholesterol”)
• AKA AAT / SERPINA1
• Member of the serine protease
inhibitor (SERPIN) family of
protease inhibitors
• ☛ Inhibits excessive protease
α1-antitrypsin activity
(α1- o ☛ Neutrophils or
antiproteinase) immunoglobulins
degranulate and release
proteases like trypsin and
elastase
• Serves as a protective screen that
prevents alveolar wall destruction
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES
• Made by the liver
• Have anti-inflammatory and
immune-regulatory activities
• Protects lungs/liver from
proteases (trypsin & elastance)
released by leukocytes
o ☛ Neutralize proteases
when inciting agents are
already eliminated
• Gene mutation – deficiency of
a1-antitrypsin
• Cigarette smoke – oxidation of
Met358
o Both gene mutation and
cigarette smoking can lead
to the loss of its inhibitory
effect → result into
excessive proteolytic activity
that can damage the lung or
the liver tissues → leading to
loss of alveolar elasticity and
increase the risk to develop
emphysema or destroy the
liver and cause cirrhosis
o Destroys protease-binding
capacity of a1-antitrypsin
• Absence/defect – increases
protease activity that can destroy
normal lung/liver tissues =
emphysema (loss of alveolar
elasticity) / cirrhosis
Figure 9.5.2-1. α1-antitrypsin
(Instructor’s slide)
• a1-acid glycoprotein
(orosomucoid)
o Bind to heparin, serotonin,
steroids, & histamine
• Retinol-binding protein
o Transports retinol secreted
by the liver to meet the
Other α1-
vitamin A requirements of
globulin
tissues
• Transcortin (Corticosteroid-
binding globulin)
o Major transporter of
glucocorticoids and
progestins
• Thyroxin-binding globulin
BIOCHEMISTRY
o One of the 3 major
transporters of thyroid
hormones (T4 & T3)
o ☛ Other 2 transporters:
transthyretin and albumin
ALPHA-2 GLOBULINS
• Carries 90% of copper in the
plasma
• “Heavenly-blue color”
• Functions:
o Aids in maintaining copper
homeostasis/transport
o Ferroxidase (Fe2+ → Fe3+)
▪ ☛ Oxidizes ferrous to
ferric iron to permit iron
binding and transport by
transferrin
Ceruloplasmin
Figure 9.5.2-2. Ferroxidase Activity of
Ceruloplasmin
(Instructor’s slide)
• ☛ Absence of this globulin can
lead to the following conditions:
o Aceruloplasminemia –
deposits iron in organs
o Wilson’s disease – deposits
copper in brain, liver, kidneys,
and cornea
• Largest plasma protein that
includes complement C3 and C4
• Functions:
o Panproteinase inhibitor
▪ Binds to any proteinases
and the complex are
cleared from the plasma
▪ ☛ Virtually capture and
α2- clear any proteinases in
macroglobulin the circulation
o Regulation of fibrinolysis,
coagulation, & complement
activation
o Binds to cytokines and 10% of
zinc and transports them to
tissues
o Plays a role in the
pathogenesis of Alzheimer’s
33
SHIFT 3| LESSON 1 | BLOOD GENERALITIES
disease, Parkinson’s disease,
& prostate cancer
• Forms a tight non-covalent
complex with free hemoglobin
released from hemolysis of red
blood cells
• Complex that is too large to be
filtered through the glomerulus
thus preventing loss of free iron in
the kidneys
• Protects the kidneys from oxidative
damage by precipitated Hgb
• Removed by the RES
o ☛ Hgb-Haptoglobin complex
is removed by macrophages
in the Reticuloendothelial
system
• Low levels seen in hemolytic
anemia
Haptoglobulin o ☛ Can result to an RBC being
destroyed more quickly than
they are produced
Figure 9.5.2-3. Haptoglobulin
(Instructor’s slide)
• ☛ VLDLs
Pre-β
lipoprotein • ☛ Major carriers of
synthesized from the liver
9.5.3. Beta globulins
BETA GLOBULINS
• Transports iron to site where it is
required
• Carries iron in its Fe3+ state
• ☛ This binding is important
Transferrin because it protects against the
toxic effects of iron in the
circulation by keeping it in its
lesser active ferric state during
transport
• Component of the Major
β2-
Histocompatibility Complex
microglobulin
(MHC) Class I molecule
• ☛ Known as the “LDL” or “bad
β-lipoprotein
cholesterol”
BIOCHEMISTRY
o It delivers cholesterol to the
tissues
• Promote complement binding &
phagocytosis of macrophages
C-reactive • ☛ Assist in complement binding of
proteins foreign and damaged cells and
enhances phagocytosis of
macrophages
• ☛ Almost similar to haptoglobin
because it also prevents iron loss
in the kidney
o The only difference is that this
Hemopexin protein binds & transport free
heme (instead of
hemoglobin) to the liver for
break down and iron
recovery
CLINICAL SIGNIFICANCE OF BETA GLOBULINS
Hemopexin • Prevent Heme-mediated
oxidative stress and iron loss
C-reactive • Acute phase reactants
proteins • Biomarker of tissue injury,
infection & inflammation
• Used to detect or monitor
significant inflammation in acute
conditions like when having
severe COVID-19 infection
Transferrin • Increase in pregnant and iron
deficient individuals
• Low in liver disease and
hemolytic anemia
Beta-2 • Tumor marker for cancers like
microglobulin multiple myeloma and lymphoma
• Form aggregates that pose a
problem during kidney dialysis
Beta- • Its elevation is associated with
lipoprotein increased risk of heart disease
and atherosclerosis
TAGs
✂ LECTURETTE/KAHOOT (LEAPNotes ‘25)
Which globulins are considered as proteinase
inhibitors to limit damaging effects of proteases to
normal tissues?
A. Alpha 2 microglobulin
B. Alpha 1 antitrypsin
C. Beta 2 microglobulin
D. Transcortin
Answer: Alpha 2 microglobulin; Alpha 1 antitrypsin
Rationale:
• Alpha 2 microglobulin is also known as a
panproteinase inhibitor.
• Proteinase inhibitors are important since they inhibit
the activity of proteinases once they have already
removed the pathogens. If not removed early,
proteinases can destroy normal tissue, especially in
the lungs and liver.
Clinical importance of C-reactive proteins:
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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

A. Absence can predispose to develop • Although haptoglobin and hemopexin, are different in
emphysema terms of type or fraction of globulin, they both bind to
B. Tumor marker for multiple myeloma & either hemoglobin or heme.
lymphoma • This binding is available so that it can bring the
C. Tumor marker for hepatocellular CA heme/hemoglobin back to the liver for resynthesis. It
D. Biomarker of acute tissue injury, infection, & can be used to resynthesize the iron in the bone
marrow instead of being lost in the kidneys.
inflammation
• Main transporter of zinc: albumin, microglobulin,
transferrin
Answer: D
Rationale: • Ceruloplasmin and Hephaestin: convert iron to its
oxidized state and bind to transferrin for recycling
• CRP are biomarkers for acute tissue injury, infection,
& inflammation. • Alpha 1 antitrypsin: control excess proteinase
activity that can damage healthy tissues
• This is why for hospitalized people, one of the blood
tests requested is the CRP Which globulin are considered as proteinase
• Absence can predispose to develop emphysema inhibitors to limit damaging effects of proteases to
[alpha-1 antitrypsin] normal tissues? (1 or more possible answer/s)
• Tumor marker for multiple myeloma & lymphoma
[beta-2 microglobulins]
• Tumor marker for hepatocellular CA / A. Alpha 2 macroglobulin
keratoblastomas [Alpha1-fetoprotein] B. Alpha 1 antitrypsin
Has a ferroxidase activity which oxidizes Fe+2 to C. Beta 2 microglobulin
Fe+3 to permit iron binding and transport by D. Transcortin
transferrin:
A. Haptoglobin Answer: A & B
B. Macroglobulin
C. Hemopexin Rationale:
D. Ceruloplasmin • Proteinases are released once inflammatory
processes help in the inflammation response to
remove/digest invading pathogens. If you don’t
Answer: D remove them early, they can destroy tissues in the
Rationale:
lung and liver. Therefore, alpha 2 macroglobulin and
• Ceruloplasmin, together with Hephaestin has alpha 1 antitrypsin are inhibitors of proteinases once
ferroxidase activity they have already removed the pathogens.
• The importance of this is that it keeps iron in the ferric
state so that it can bind to transferrin during transport
What globin protein fraction does pre-beta 9.6. PLASMA PROTEINS IN DIAGNOSIS OF DISEASES (JLN)
lipoproteins belong to? • Electrophoresis – separates plasma proteins
A. Alpha 1 according to their net negative charge and
B. Beta movement towards the anode
C. Alpha 2 o ☛ Commonly used in diagnosis of
D. Gamma diseases affecting plasma proteins
• Vertical axis or intensity of band correlates with the
Answer: C plasma protein concentration

Rationale:
• Pre-beta lipoproteins are also known as VLDL. VLDL
belongs to Alpha 2 globulins.
• HDL are alpha 1 lipoproteins.
• LDL are beta lipoproteins.
Why are haptoglobin and hemopexin important in
the blood circulation?
A. Control excess proteinase activity that can
damage healthy tissues
B. Bine extracorpuscular hemoglobin/heme to
avoid iron loss in the kidney
C. Convert iron to its oxidized state and bind to
transferrin for recycling
D. Transport zinc in the plasma for proper function
of the immune system

Answer: B
Rationale: Figure Explanation

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SHIFT 3| LESSON 1 | BLOOD GENERALITIES

• Plasma proteins are mostly Hepatic Cirrhosis

Above: negatively charged so they tend (Polyclonal Gammopathy)
Normal to move towards the anode.
electrophoretic graph • Broad elevation of
pattern of various gamma globulins with
plasma proteins at
reduction in albumin
pH=8.6.
concentration
Below: • Albumin traveled the farthest
Pattern in agarose Paraprotein (Monoclonal
towards the anode and the
gel. Gammopathy)
gamma globulin the slowest.
• Height of the band along the
• Ex. Multiple myeloma or
vertical axis or the intensity in
amyloidosis
gel electrophoresis show the
• Sharp rise in gamma
relative concentration of each
globulin band
plasma protein fraction with
albumin proving to be the most
abundant among them.

Nephrotic Syndrome
9.6.1. Plasma Protein Electrophoresis in the Diagnosis of
Diseases
• DEC albumin
• INC alpha-2 macroglobulin
Many diseases alter the plasma protein levels in the and pre-beta lipoprotein in
blood and can be detected using electrophoresis the alpha-2 globulin band
Normal pattern in
Electrophoresis of plasma
protein levels Protein-losing enteropathy

• Resulting from intestinal

problems like inflammatory
bowel disease, lymphatic
obstruction and alterations
Immediate Response
in capillary permeability
Pattern
• Marked DEC in albumin
and gamma globulin
• Immediate response to
fractions
stress or inflammation
• INC alpha-2 globulins
• Caused by infection, injury
or surgical trauma → INC
haptoglobins (alpha-2 Causes of Hypogammaglobulinemia (Not Discussed)
band)
Delayed Response Primary Causes Examples
Pattern Severe combined
T cell defects immunodeficiency;
• Delayed response → INC Hyper-IgM syndrome
immunoglobulins (gamma X-linked agammaglobulinemia;
B cell defects Common variable
band)
immunodeficiency
Secondary Causes Examples
Hypogammaglobulinemia Malignancy;
Decreased Medications;
production Infection (perinatally acquired HIV);
• May result from Starvation
immunosuppressive High catabolism (very rare);
diseases causing Loss Protein-losing enteropathy;
flattening of concentration Drainage of ascites or chylothorax
of gamma globulin fraction
MONOCLONAL GAMMOPATHIES (Not Discussed)
MALIGNANT
• Plasma cell disease

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• Monoclonal gammopathy of gastroenteritis, Celiac sprue, allergic

undetermined significance protein losing enteropathy, Giardiasis or
• Multiple myeloma hookworm infection, amyloidosis,
• Smoldering multiple myeloma immunodeficiency, SLE
• AL amyloidosis
• Other rarer malignant plasma cell REFERENCES
disorders Intructor’s Slides, LeapNotes Batch 2025, Lecturette
• B-cell (usually IgM) disease
o Lymphoplasmacytic TWG TL: Y2K (1A)
lymphoma/Waldenstrom
macroglobulinemia
o Chronic lymphocytic leukemia APPENDICES
o Small lymphocytic lymphoma
o Marginal zone lymphoma
o Other indolent lymphomas (rare)
BENIGN
• Autoimmune/Inflammatory disease
• Rheumatoid arthritis, ankylosing
spondylitis
• Systemic lupus erythematosus,
scleroderma, Sjogren syndrome
• Vasculitis, polymyalgia rheumatica
• Paraprotein-associated neuropathies

PROTEIN LOSING ENTEROPATHY (Not Discussed)

3 MECHANISMS
• Mucosal disease with ulceration
• Chronic gastric ulcer
• Gastric carcinoma
• Lymphoma
• Inflammatory bowel disease
• Idiopathic ulcerative jejunoileitis
• Lymphatic obstruction
o Primary intestinal lymphangiectasia
o Secondary obstruction due to heart
disease, infection, neoplasm,
retroperitoneal fibroses, or sarcoidosis
• Alterations in mucous capillary permeability
o Menetrier’s disease
o Zollinger-Ellison syndrome
o Acute viral or eosinophilic gastroenteritis
o Celiac sprue
o Allergic protein losing enteropathy
o Giardiasis or hookworm infection
o Amyloidosis, Immunodeficiency. SLE

PROTEIN LOSING ENTEROPATHY (Not Discussed)

3 MECHANISMS
• Mucosal Disease with ulceration
o Chronic gastric ulcer, gastric carcinoma,
lymphoma, inflammatory bowel disease,
idiopathic ulcerative jejunoileitis
• Lymphatic Obstruction
o Primary intestinal lymphangiectasia,
Secondary obstruction due to heart
disease, infection, neoplasm,
retroperitoneal fibrosis, or sarcoidosis
• Alterations in mucous capillary permeability
o Menetrier’s disease, Zollinger-Ellison
syndrome, acute viral or eosinophilic
BIOCHEMISTRY 37
SHIFT 3| LESSON 1 | BLOOD GENERALITIES

NEUTROPHILS EOSINOPHILS BASOPHILS MONOCYTE LYMPHOCYTE

60-70% 2-4% 0-1% 5% 25%

12-15 μm 12-15 μm 12-15 μm 12-20 μm 6-18 μm

Few hours to few

7 hours 8-12 days 3 days May live for years
days

FREEDOM WALL

BIOCHEMISTRY 38