Isoenzymes

Isoenzymes

Isoenzymes or isozymes are multiple forms of same enzyme

that catalyze the same chemical reaction
Different chemical and physical properties:
Electrophoretic mobility
Kinetic properties
Amino acid sequence
Amino acid composition
 Lactate Dehydrogenase

 Lactate dehydrogenase (LDH) is an enzyme present in a wide variety

of organisms
 EC 1 = oxidoreductase.
 EC 1.1 = acting on the CH-OH group of the donor.
 EC 1.1.1 = With NAD or NADP as acceptor.
 EC 1.1.1.27 = L-lactate dehydrogenase.
 Molecular weight- 32 kD & it is tetramer
M (A) -muscle –chromosome 11(basic)
H (B) -heart – chromosome 12(acidic)
Lactate dehydrogenase, reversibly converts lactate to pyruvate, in
different tissues.
LDH consists of 5 iso-enzymes – LDH1,LDH2,LDH3,LDH4 &
LDH5
These isoenzymes are separated by cellulose acetate electrophoresis
at pH 8.6
Normal values:
Serum -100 -200 U/L
CSF - 7 -30 U/L
Urine - 40 -100 U/L
LDH reaction
LDH isoforms
 Clinical significance of LDH

 In normal serum, LDH2 (H3M) predominant isoenzyme & LDH5

is rarely seen.
 In myocardial infarction, LDH1(H4) levels are greater than
LDH2.
 Megaloblastic anemia (50 times upper limit of LDH 1 and LDH
2)
 Muscular dystrophy, LDH5 (M4) is increased.
 Toxic hepatitis with jaundice (10 times more LDH5)
Renal disease- tubular necrosis or pyelonephritis, pulmonary embolism
LDH 3 (massive destruction of platelets)
Total LDH is increased in neoplastic diseases.
LDH5 is increased in breast cancer, malignancies of CNS, prostatic
carcinoma.
In leukaemias, LDH2 & LDH3 levels are increased.
In malignant tumors of testis & ovaries, LDH2, LDH3 & LDH 5 levels are
increased.
In CSF:

Bacterial meningitis – LDH4 and LDH5

Viral meningitis - LDH1

Metastatic tumors - LDH5

Neonatal cases of intracranial haemorrhage associated with

seizures and hydrocephalus

Creatinine phosphokinase (CPK)
Creatine Phosphokinase (CPK)
Creatine phosphokinase isoenzymes
 Clinical significance of CK

 CPK & heart attack:

 CPK2 isoenzymes is very small, (2% of total CPK activity) &
undetectable in plasma.
 In myocardial infarction (MI), CPK2 levels are increased within 4
hrs, then falls rapidly.
 Total CPK level is elevated upto 20-folds in MI.
 CPK & Muscle diseases

 CPK level is elevated in muscular dystrophy (500-1500U/L)

 CPK level is highly elevated in crush injury, fracture & acute
cerebrovascular accidents.
 Estimation of total CPK is employed in muscular dystrophies &
CPK-MB isoenzyme is estimated in myocardial infarction.
 Atypical forms of CK

Two atypical isoforms.

1. Macro-CK (CK-macro)
Formation:

Formed by aggregation CK-MB with IgG, sometimes IgA.

Also formed by complexing CK-MM with lipoproteins.
Electrophoretically migrates between CK-MB & CK-MM.
Occurs frequently in women above 50 years.
 CK-Mi (Mitochondrial CK-Isoenzyme)

Formation:

It is present bound to the exterior surface of inner mitochondrial

membrane of muscle, liver & brain.
It exist in dimeric form & oligomeric aggregates & molecular weight
35,000
Electrophoretically, migrates towards cathode & is behind CK-MM
band.
It is not present in normal serum.
 Clinical significance

It is present in serum when there is extensive tissue damage causing

breakdown of mitochondrial & cell wall.
Its presence in serum indicates cellular damage, seen in malignancies.
Thank you