LDH & CK

BY NDARUBWEINE JOSEPH

MLS 3 LECTURE

5/03/2019
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
 LDH isoforms

Isoenzyme Composition Electrophoretic Present in Elevated in

name migration

LDH 1
Heat resistant ( H4) Fastest moving Myocardium, myocardial infarction
RBC, kidney
LDH2 Myocardium, Kidney disease,
Heat resistant (H3M1) RBC, kidney megaloblastic anemia

LDH3 (H2M2) brain Leukemia, malignancy

LDH4 (H1M3) Lung, spleen Pulmonary infarction

Heat labile

LDH5
Heat labile (M4) Slowest moving Skeletal Skeletal muscle and liver
Inhibited by muscle, Liver diseases
urea
 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
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)
It catalyses creatine to creatine phosphate.
 Normal serum value:
 15-100 U/L for males & 10-80 U/L for females.
CPK consists of 3 isoenzymes.
Each isoenzyme of CK is a dimer;
Molecular weight of 40 kD.
The subunits are called B for brain (chromosome -
14) & M for muscle (chromosome -19)
 Creatine
Phosphokinase (CPK)

It is an important enzyme in energy

metabolism.
Immediate source of ATP in contracting
Three Iso-enzymes are separated by electrophoresis.

CPK-1 (also called CPK-BB) is found mostly in the

brain & lungs.

CPK-2 (also called CPK-MB) is found mostly in the

heart.

CPK-3 (also called CPK-MM) is found mostly in

 Creatine phosphokinase
isoenzymes

ISOENZYMES SUB- TISSUE % IN SERUM

UNIT

CK1 BB Brain 1
Fast moving

CK2 MB Heart 5
2% of total

CK3
Slow moving MM Skeletal muscle 80
 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-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