Chapter 4

Hemoglobin Function and

Principles of Hemolysis

Copyright ©2019 F.A. Davis Company

 Hemoglobin Synthesis
 The function of the R B C’s is to produce,
package, protect, and transport hemoglobin
among various tissues
 Hemoglobin
• Responsible for carrying oxygen (oxyhemoglobin)
• Pulls C O2 away from tissues
• Keep balanced p H between the blood

Copyright ©2019 F.A. Davis Company

 Red Cell Composition
 Heme
• 4 iron atoms iron2+ (linked through histidine a.a)
• Iron surrounded by protoporphyrin ring (multi-
step enzymatic procedure)
• ferroprotoporphyrin
 Globin
• 2 pairs of globin chains made up of amino acids
• Alpha and beta chains

Copyright ©2019 F.A. Davis Company

 Red Cell Composition
 Hemoglobin
• 1o structure = a.a. sequence
• 2o structure = helices and non-helices
• 3o structure = globular arrangements of individual
chains
• 4o structure = tetramer formation of complete hgb
 2,3-Diphosphoglycerate (2,3-D P G)
• Produced via Embden-Meyerhof pathway
• Related to oxygen affinity of hemoglobin

Copyright ©2019 F.A. Davis Company

 Hemoglobin Molecule

Copyright ©2019 F.A. Davis Company

 Genetics and Chain Formation of
Hemoglobin
 Chromosome 11
• Genes for epsilon, beta, gamma G and A, delta
chains
 Chromosome 16
• Alpha and zeta genes
 3 types of hemoglobin in red cell production
• Embryonic hemoglobin
• Fetal hemoglobin
• Adult hemoglobin

Copyright ©2019 F.A. Davis Company

 Hemoglobin
 Embryonic hemoglobin
• Hemoglobin Gower one (ζ2, ε2)
• Hemoglobin Gower two (α2 ε2)
• Hemoglobin Portland (ζ2 γ2)
 Fetal hemoglobin (3 months of fetal
development)
• Hemoglobin F (α2 γ2) – up to 90%

Copyright ©2019 F.A. Davis Company

 Hemoglobin (continued)
 Adult hemoglobin (3 to 6 months after
delivery)
• Hemoglobin A (α2 β2): 95% to 98%
• Hemoglobin A2 (α2 δ2): 3% to 5%
• Hemoglobin F (α2 γ2): less than 2%

 Table 4.1

Copyright ©2019 F.A. Davis Company

 Oxygen Dissociation (O D) Curve
 The binding and release (reversible) of
oxygen from the hemoglobin molecule
 1g hgb = 1.34mL oxygen
 bind oxygen = oxygen-rich
 release oxygen = oxygen-poor
 binding causes allosteric alterations
• tense
• relaxed

Copyright ©2019 F.A. Davis Company

 Oxygen Dissociation (O D) Curve
 OD curve
• Sigmoid shape (“S” shape)
• Partial pressure of O2 (PO2) = P50 value
• ↑ bound oxygen when ↑ PO2
• Lungs: PO2 is 100 millimeters of mercury = 97%
Hemoglobin saturation with oxygen
• Circulation: PO2 is 40 millimeters of mercury =
75% hemoglobin saturation with oxygen

Copyright ©2019 F.A. Davis Company

 Oxygen Dissociation (O D) Curve
(continued)

Copyright ©2019 F.A. Davis Company

 O D Curve Changes
 Shift to right
• Less attraction to oxygen (release)
• At 40 millimeters of mercury, hemoglobin is 50%
more saturated but willing to give up 50% of
oxygen to tissue if needed
• Lower affinity
‒ Anemia
‒ Acidosis (decreased p H)
‒ Increased 2,3-D P G
‒ Elevated temperature (fever)

Copyright ©2019 F.A. Davis Company

 O D Curve Changes (continued)
 Shift to left
• More attraction for oxygen
• At 40 millimeters of mercury, hemoglobin is 75%
more saturated but willing to release only 12% to
tissues
• Higher affinity
‒ Presence of abnormal hemoglobins (high affinity)
‒ Alkalosis (increased p H)
‒ ↓ 2,3-D P G, body temp
‒ Multiple transfusions of stored blood where 2,3-D P G
is depleted

Copyright ©2019 F.A. Davis Company

 CO2 Transport
 secondary function
 diffuses into RBC
 complex interactions and reactions of
carbonic acid (H2CO3), bicarbonate (HCO3-),
chloride, H+ ions and water
 CO2 from tissues  transported by
hemoglobin as H+  combines with
bicarbonate to form carbonic acid  H2O
and CO2  expelled from lungs

Copyright ©2019 F.A. Davis Company

 Abnormal Hemoglobins
 Methemoglobin (MetHb)
• Iron in iron3+ state; Not capable of binding oxygen
• If greater than 10%, individuals appear cyanotic
(blue color)
• dyspnea, headache, change in mental status
• Can be induced by aniline drugs or hemoglobin M
(autosomal dominant)
• Avoid offending oxidizing agent
• Treatment: IV Methylene Blue or exchange
transfusions may be required

Copyright ©2019 F.A. Davis Company

 Abnormal Hemoglobins
 Carboxyhemoglobin (COHb)
• High affinity for carbon monoxide
• Oxygen cannot be delivered to tissues
• Can lead to carbon monoxide poisoning
• Increased in smokers and certain industrial workers
• Headache, dizziness, disorientation
• Blood appears cherry red
• COHb level = greater than 3-10%
• Treatment = oxygen therapy

Copyright ©2019 F.A. Davis Company

 Abnormal Hemoglobins (continued)
 Sulfhemoglobins
• Affinity for oxygen is lower
• Formed through exposure to sulfonamides or
sulfa-containing drugs
• Persists for life of cell
• Avoid offending agent

Copyright ©2019 F.A. Davis Company

 Hemolysis
 Cells age  Destruction of R B C’s
 Interruption of cholesterol and phospholipid
management
 Loss of selective permeability
 Red cell becomes spheroidal
 SA:V decreases
 Triggered when ATP generation decreases
 Enzymes are destroyed

Copyright ©2019 F.A. Davis Company

 Hemolysis
 Extravascular 90%
• available energy stores are reduced
• R E S system: spleen, liver, lymph nodes, bone
marrow
• Releases heme and globin contents to be
recycled
• Globin, Heme
• Iron  transferrin  BM

Copyright ©2019 F.A. Davis Company

 Figure 4.4

Copyright ©2019 F.A. Davis Company

 Extravascular Hemolysis
 ↓ Hemoglobin, H C T, R B C count
 ↑ Reticulocyte count (if bone marrow working)
• Polychromasia in peripheral smear
 ↑ Serum bilirubin
 ↓ Haptoglobin
↑LDH
 Spherocytes
 Urine: ↑ urobilinogen

Copyright ©2019 F.A. Davis Company

 Hemolysis
 Intravascular
• Lysed directly into blood vessels
• free hemoglobin = haptoglobin
• during complement activation (ABO transfusion
reactions)

Copyright ©2019 F.A. Davis Company

 Intravascular Hemolysis

 ↓ Hemoglobin, H C  Hemoglobinuria (red

T, R B C count urine)
 ↑ Serum bilirubin  Possible ↑
 ↓ Serum reticulocytes
haptoglobin ↑LDH
 Hemoglobinemia  Schistocytes
(free hemoglobin):  Urine: ↑
plasma may appear urobilinogen
red or pink-tinged

Copyright ©2019 F.A. Davis Company

 Hemolysis Response
 Bone Marrow
• erythroid hyperplasia
• M:E ratio ~1:2
• early release of erythroid precursors

 Review Table 4.2

Copyright ©2019 F.A. Davis Company

 Classifications Relevant to the Hemolytic
Anemias

Hemolytic Anemias Classified by Intrinsic or Extrinsic Defects (Modified List)

Intrinsic Red Blood Cell
Defects Leading to Hemolysis
Hemoglobinopathies: structural
and synthetic
Red blood cell membrane defects
Red blood cell enzyme defects
Stem cell defects
Extrinsic Defects Leading to Hemolysis
Autoimmune hemolytic anemia
Parasitic infection
Microangiopathic hemolytic anemia
Environmental agents, including venoms
and chemical agents

Copyright ©2019 F.A. Davis Company

 Homework
 Read Chapter 4

 Test Next Week!

• Cell descriptions and information still relevant
• Safety

Copyright ©2019 F.A. Davis Company