Introduction to Clinical Enzymology

(Diagnostic Plasma Enzymes)

Chidum Ezenwaka
Professor of Chemical Pathology

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 Objectives – Learning outcome
At the end of this lecture, students should know:
 The meaning of diagnostic enzymes

 Characteristics of enzymes

 Some enzymes used for disease diagnosis

 Sources and uses of some diagnostic

plasma enzymes

 What causes decrease or increase in plasma

enzymes

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Characteristics of enzymes
 Enzymes are proteins that act as biological
catalysts, altering the rate, and providing a means
of regulating metabolic reactions

 most easily measured in terms of biological

activity rather than their mass

 fairly constant in healthy individuals, some

increase after muscular exercise (CK) or after a
meal (intestinal isoenzymes of ALP)

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Characteristics of enzymes cont’d
 Most diagnostic enzymes have half-life of 10
hours to >5 days

 enzyme activity is usually measured -

concentrations can also be measured

 unit of measurement of enzyme is International

Unit (IU): defined as ‘that amount of enzyme
which under given assay conditions will catalyse
the conversion of 1mol of substance per
minute’.

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Causes of enzyme increase in plasma:

 severe damage to cell – ischaemia or toxic

substances mostly affecting cytoplasmic enzymes
(e.g. ALT)

 increased rate of cell turnover – occurs during

periods of active growth (physiological e.g. ALP),
tissue repair or in association with several forms of
malignant disease (cancer)

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Causes of enzyme increase in plasma cont’d
 increased concentration of enzymes within cells
– enzymes whose synthesis is induced by disease
or drugs e.g. GT synthesis is induced by ethanol

 duct obstruction – occurs in enzymes that are

normally present in exocrine secretions. This is
regurgitated into the blood if the normal route
of outflow is obstructed – e.g. ALP

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 Causes of decreased levels of plasma
enzymes:

 reduced cell population (severe liver failure)

 genetic deficiencies (e.g.

hypophosphatasemia)

 reduced activity (organophosphate poisoning

of pseudocholinesterase)

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 Factors affecting the numerical values of
enzyme results:
 nature and concentration of substrate

 reaction temperature

 pH (H+ concentration)

 type of buffer

 nature and concentration of co-factors etc.

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 Questions to consider before requesting to
do plasma enzyme tests

 which organ/tissue is damaged? - test

specificity

 what is the extent of damage? - test sensitivity

 Is the plasma enzyme activity changing during

the course of the disease?

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 Choose a biomarker - Year 3 clerkship!!

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 Isoenzymes

 These are proteins that possess similar

catalytic activity but show genetically
determined differences in their structure and
certain other properties such as
electrophoretic mobility, stability to heat etc.

 Isoenzymes aid in localizing the tissue of

origin in an increased plasma enzyme
activity.

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 Common diagnostic plasma enzymes
1. Amylase:
Source: pancreas, salivary gland, lungs,
fallopian tubes, ovary and
prostrate.
Half life: ~ 12 h

Use: diagnosis of acute pancreatitis,

alcoholism, biliary tract disease,
hyperlipidaemia, truma etc.

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 Common diagnostic plasma enzymes cont’d

2. Creatine Kinase
- has 2 subunits of 2 polypeptides designated M or B.
These subunits combine to form 3 isoenzymes MM,
MB and BB.

Source: MM skeletal and cardiac muscle

MB cardiac muscle
BB brain (present in CSF)

Use: diagnosis of cardiac diseases:- myocardial

infarction (MI), myocarditis, skeletal muscle
damage etc.

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 Common diagnostic plasma enzymes cont’d

3. Aspartate aminotransferase (AST)

Source: heart, liver, skeletal muscle,
pancreas, kidney,
erythrocytes.
Use: diagnosis of myocardial
disease, liver disease, skeletal
muscle disease etc.

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 Common diagnostic plasma enzymes cont’d

4. Alanine aminotransferase (ALT)

Source as above but liver
dominant.
Use diagnosis of hepatocellular
damage, more liver specific
than AST. Raised in
cardiac failure, muscle
disease etc.

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 Common diagnostic plasma enzymes cont’d
5. Lactate dehydrogenase (LD)
- LD is a tetramer with 2 subunit H (heart) and M
(muscle) & 5 isoenzymes - LD1, LD2, LD3, LD4, LD5

LD1 & LD2 - heart

LD5 - liver

Source: widespread cytosolic enzyme,

Major: heart, skeletal muscle, liver, kidney,
erythrocytes etc.
Use: diagnosis of myocardial disease, liver
disease, skeletal muscle disease etc.

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 Common diagnostic plasma enzymes cont’d

6. Alkaline phosphate (ALP)

- has 4-5 known but poorly characterised
isoenzymes

Source: bone (osteoblasts), liver, intestine,

kidney, placenta.

Use: diagnosis of liver disease

(cholestasis), bone disease
(osteoblastic activity) e.g.
Paget’s disease, hyperparathyroidism,
malignancy, bone tumours, liver
tumours etc.

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 Common diagnostic plasma enzymes cont’d

7. Gamma glutamyltransferase (GT)

- synthesis is induced by alcohol and
drugs such as barbiturates, phenytoin
etc.
Source: liver (not in bone)

Use: differential diagnosis of liver

disease (alcoholic cirrhosis).

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 Common diagnostic plasma enzymes cont’d

8. 5’Nucleotidase
- technically difficult to measure, not
sensitive and not widely used presently.

Source: - in a variety of tissues.

Use: - diagnosis of hepatobiliary

disorders
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 Common diagnostic plasma enzymes cont’d
9. Acid phosphatase (ACP)
- up to 14 bands of isoenzymes are seen on
electrophoresis.

Main source: prostate (100-400x others)

Other sources:erythrocytes, white cells, platelets,
Gaucher’s cells.

Use: diagnosis of prostatic carcinoma, metastatic

disease of bone, Paget’s disease,
thrombocytopenia purpura etc.

NB: levels are raised in plasma after rectal examination

or haemolysis.

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 Common diagnostic plasma enzymes cont’d
10. Cholinesterase
- 2 principal cholinesterase exist:
• pseudocholinesterase - synthesised in the liver
• acetylcholinesterase (true cholinesterase) - present
at nerve endings and in the erythrocytes but not in
plasma

Source: plasma, liver, nervous tissue, red cells.

Use: diagnosis of patients with scoline apnoea

and organophosphorous insecticide
poisoning.

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 End

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