Professional Documents
Culture Documents
Notes in Enzymology
Notes in Enzymology
Notes in Enzymology
Tissue Source
ALT is distributed in many tissues, with comparatively high concentrations in the liver. It is
considered the more liver-specific enzyme of the transferases.
Diagnostic Significance
Confined mainly to evaluation of hepatic disorders.
In acute inflammatory conditions of the liver, ALT elevations are frequently higher than those
of AST and tend to remain elevated longer as a result of the longer half-life of ALT in serum
(16 and 24 hours, respectfully).
Source of Error
ALT is stable for 3 to 4 days at 4°C. It is relatively unaffected by hemolysis.
2. ALKALINE PHOSPHATASE
Alkaline phosphatase (ALP) belongs to a group of enzymes that catalyze the hydrolysis of
various phosphomonoesters at an alkaline pH.
ALP functions to liberate inorganic phosphate from an organic phosphate ester with the
concomitant production of an alcohol.
The optimal pH for the reaction is 9.0 to 10.0, but optimal pH varies with the substrate used.
The enzyme requires Mg2+ as an activator.
Tissue Source
ALP activity is present on cell surfaces in most human tissue. The highest concentrations are
found in the intestine, liver, bone, spleen, placenta, and kidney.
In the liver, the enzyme is located on both sinusoidal and bile canalicular membranes;
activity in bone is confined to the osteoblasts, those cells involved in the production of bone
matrix.
Diagnostic Significance
Elevations of ALP are of most diagnostic significance in the evaluation of hepatobiliary and
bone disorders.
In biliary tract obstruction, ALP levels range from 3 to 10 times ULN. Increases are primarily a
result of increased synthesis of the enzyme induced by cholestasis.
Elevated ALP levels may be observed in various bone disorders. Perhaps the highest
elevations of ALP activity occur in Paget’s disease (osteitis deformans).
In normal pregnancy, increased ALP activity, averaging approximately 1 1⁄2 times ULN, can
be detected between weeks 16 and 20. ALP activity increases and persists until the onset of
labor. Activity then returns to normal within 3 to 6 days.
ALP levels are significantly decreased in the inherited condition of hypophosphatasia.
Subnormal activity is a result of the absence of the bone isoenzyme and results in inadequate
bone calcification.
3. Y-Glutamyltransferase
y-Glutamyltransferase (GGT) is an enzyme involved in the transfer of the y-glutamyl residue
from y-glutamyl peptides to amino acids, H2O, and other small peptides. In most biologic
systems, glutathione serves as the y-glutamyl donor.
The specific physiologic function of GGT has not been clearly established, but it is suggested
that GGT is involved in peptide and protein synthesis, regulation of tissue glutathione levels,
and the transport of amino acids across cell membranes.
Tissue Source
GGT activity is found primarily in tissue of the kidney, brain, prostate, pancreas, and liver.
Clinical applications of assay, however, are confined mainly to evaluation of liver and biliary
system disorders.
Diagnostic Significance
In the liver, GGT is located in the canaliculi of the hepatic cells and particularly in the
epithelial cells lining the biliary ductules.
Because of these locations, GGT is elevated in virtually all hepatobiliary disorders, making it
one of the most sensitive of enzyme assays in these conditions. Higher elevations are
generally observed in biliary tract obstruction.
Because of the effects of alcohol on GGT activity, elevated GGT levels may indicate
alcoholism, particularly chronic alcoholism.
GGT assays are useful in monitoring the effects of abstention from alcohol and are used as
such by alcohol treatment centers. Levels usually return to normal within 2 to 3 weeks after
cessation but can rise again if alcohol consumption is resumed.
GGT activity is useful in differentiating the source of an elevated ALP level because GGT
levels are normal in skeletal disorders and during pregnancy.
Source of Error
GGT activity is stable, with no loss of activity for 1 week at 4°C. Hemolysis does not interfere
with GGT levels because the enzyme is lacking in erythrocytes.
Reference Range
GGT: male, 6–45 U/L (37°C); female, 5–30 U/L (37°C).
Values are lower in females, presumably because of suppression of enzyme activity resulting
from estrogenic or progestational hormones.
CARDIAC ENZYMES
CK-MB
AST
LD (LDH) 1 & 2
LD/HBD ratio
MI Biomarkers
1. CREATININE KINASE
Tissue Source
CK is widely distributed in tissue, with highest activities found in skeletal muscle, heart
muscle, and brain tissue.
CK is present in much smaller quantities in other tissue sources, including the bladder,
placenta, gastrointestinal tract, thyroid, uterus, kidney, lung, prostate, spleen, liver, and
pancreas.
Has 3 isoenzymes:
CK-BB (CK1)= Brain
CK-MB (CK2)= Heart, muscle
CK-MM (CK3)= Muscle, heart
Cardiac muscle – CK-MM and CK-MB.
Skeletal muscle – CK-MM
Brain, GI, colon, prostate, uterus = CK=BB
Trauma to skeletal muscle causes increase in total CK and MB isoenzyme, but % activity
of MB is <3% (>6% in MI)
2. Aspartate Aminotransferase
It is commonly referred to as a transaminase and is involved in the transfer of an amino group
between aspartate and a-keto acids.
The older terminology, serum glutamic-oxaloacetic transaminase (SGOT, or GOT), may also
be used.
Pyridoxal phosphate functions as a coenzyme
Tissue Source
AST is widely distributed in human tissue.
The highest concentrations are found in cardiac tissue, liver, and skeletal muscle.
With smaller amounts found in the kidney, pancreas, and erythrocytes.
Diagnostic Significance
The clinical use of AST is limited mainly to the evaluation of hepatocellular disorders and
skeletal muscle involvement.
In AMI, AST levels begin to rise within 6 to 8 hours, peak at 24 hours, and generally return to
normal within 5 days. However, because of the wide tissue distribution, AST levels are not
useful in the diagnosis of AMI.
Source of Error/C
Hemolysis should be avoided because it can dramatically increase serum AST concentration.
AST activity is stable in serum for 3 to 4 days at refrigerated temperatures.
CLINICAL SIGNIFICANCE
Increase in:
Anemia (pernicious, hemolytic, megaloblastic)
Myocardial infarction (MI)
Muscle trauma
Renal infarct
MYOGLOBIN
Major protein responsible for oxygen supply of striated muscle
TROPONIN
the troponin complex is a component of the thin filament of striated muscle linked to actin
Three Subunits:
Troponin I: an inhibitory subunit
Troponin T: tropomyosin-binding subunit
Troponin C: Calcium-binding subunit
Elevation after 2-4 hours 3-4 hours 3-12 hours 4-8 hours 6-8 hours 8-10 hours
chest pain (MI)
Peak activity 6-10 hours 48 hours 12-24 hours 12 - 24 hours 24-48 hours 72 hours
Duration of 2-5 days 2-5 days 5-10 days 2-3 days 4 days 10 days
elevation
Sensitivity Sensitive but More sensitive More sensitive Not entirely Not sensitive, Insensitive,
not specific and specific and specific specific for AMI not specific Nonspecific
than CK-MB than CK-MB
Usefulness Negative Eliminates Eliminates Gold standard Detect Detect
predictable need for LD need for LD infarction > 3 infarction > 5
marker isoenzyme isoenzyme days prior to days prior to
testing testing
1. Amylase
found in the SALIVARY GLANDS and PANCREAS
Breaks down starch to simple sugars
Substrate: starch
starch/ amylum maltose glucose
Substrates:
o Pancreatic AMS: diastase
o Salivary AMS: ptyalin
Serum AMS is usually pancreatic in origin
o microAMS: unbound, free. 50,000 dal. AMS found in urine
o macroAMS: bound to IgG, IgA. High MW. Measured in serum
Methods
METHODS PRINCIPLES
2. Lipase
Breaks down Triglyceride into fatty acids and glycerol
Significance: Acute Pancreatitis
Substrate: Olive Oil
End Product: Fatty Acids
Methods:
o Cherry-Crandall
o Sigma-Tietz
o Titration
1. ACP (pACp)
Optimum pH: 5
Very Labile, Add 5M acetate buffer or citrate tablet to preserve
ACP are found in Prostate, RBC, bone, liver, kidneys, platelets
Substrate: organic Phosphate such as β-glyceroPO4 and ρ-nitrophenylPO4
Method
Chemical Inhibition Test
If Total ACP is normal: Stop test
If elevated: suggestive of prostatic CA
do p-ACP by Chemical Inhibition Test
Cu++ Tartrate C4H4O6-2
s
2. PSA (Prostate-Specific Antigen)
Member of the kallikrein family of serine proteases uniquely produced form the epithelial cells of
the prostate gland
Most useful TM for Prostate Cancer
Drawback: weak in distinguishing prostate CA from nonmalignant prostate lesions
Reference range: 0-4ng/mL