Isoenzymes

Isoenzymes
Multiple forms of enzymes (isoenzymes) occur
which have similar catalytic activities but
different structures.
Different isoenzymes are often organ-specific
and their determination may improve the
specificity of enzyme tests.
The heterogeneity of some isoenzymes is due to
different protein subunits which are coded for by
separate genes.

Isoenzyme: Differences
Charge: Can be separated by electrophoresis
Stability to heat denaturation
Reaction to chemical inhibitors
Affinity for substrates or coenzymes
The most common ones: LDH, CK, ALK and ACP

Reasons for isoenzyme

Synthesized from different genes (malate
dehydrogenase in cytosol versus in mitochondria)
Oligomeric forms of more than one type of
subunits (lactate dehydrogenase)
Different carbohydrate content (alkaline
phosphatase)

Lactate dehydrogenase (LDH)

5 isoenzymes, LDH1 LDH5

Tetramer
M subunits (M for muscle), basic
H subunits (H for heart), acidic

Different catalytic activities

Used as the marker for disease diagnosis

Lactate dehydrogenase (LDH)

isoenzymes with slightly different subunits

Isoenzyme Composition Composition Present in

name

Elevated in

LDH1

( H4)

HHHH

Myocardium myocardial
, RBC
infarction

LDH2

(H3M1)

HHHM

Myocardium
, RBC

LDH3

(H2M2)

HHMM

Kidney,
Skeletal
muscle

LDH4

(H1M3)

HMMM

Kidney,
Skeletal
muscle

LDH5

(M4)

MMMM

Skeletal
muscle,
Liver

Skeletal
muscle and
liver diseases

Creatine phosphokinase
(CK, CPK)

Creatine phosphokinase (CK, CPK)

CK occurs in 3 isoenzymes, each is a dimer
composed of 2 subunits [M (muscle) or B
(brain)]: CK1 = BB from brain, CK2 = MB from
the heartand CK3 = MM from skeletal muscle.
Each CK isozyme shows a characteristic
electrophoretic mobility.

Creatine phosphokinase (CK, CPK)

CPK2 is undetectable (<2%) in serum for healthy
individuals, and elevated to 20% in the first 6-18
hrs after myocardial infarction.
Used as a earliest reliable indicator of
myocardial infarction.

Creatine phosphokinase (CK, CPK)

Isoenzyme
Composition Present in
name

Elevated in

CK-1

CNS diseases

CK-2

CK-3

BB

Brain

MB

Acute
Myocardium/
myocardial
Heart
infarction

MM

Skeletal
muscle,
Myocardium

Alkaline phosphatase (ALP)

Maximal activity at a high pH 9.0-10.5

Widely distributed throughout the body

High levels are seen is liver, bone, placenta

and intestine and useful to assess
hepatobiliary and bone diseases

ALP Isoenzymes
Humans and most other mammals contain the
following ALP isoenzymes:
ALPI intestinal
ALPL tissue non-specific (liver/bone/kidney)
ALPP placental (Regan isozyme)

All mammalian ALP isoenzymes except placental

are inhibited by homoarginine.
Heating for ~2 hours at 65C inactivates most
isoenzymes except placental isoforms.

Pyruvate Dehydrogenase
Complex (PDC)
(Multienzyme Complex)

Pyruvate Dehydrogenase Complex

(PDC)
large complex containing many
copies of each of 3 enzymes, E1,
E2, & E3.
The inner core of mammalian PDC
is consisting of 60 copies of E2.
At the periphery of the complex are:

30 copies of E1 (itself a tetramer with subunits a2b2).

12 copies of E3 (a homodimer), plus 12 copies of an E3
binding protein that links E3 to E2.

Pyruvate Dehydrogenase Subunits

Enzyme

Abbreviated

Prosthetic Group

Pyruvate
Dehydrogenase

E1

Thiamine
pyrophosphate (TPP)

Dihydrolipoyl
Transacetylase

E2

Lipoamide

Dihydrolipoyl
Dehydrogenase

E3

FAD

DNA polymerase (Multifunctional

Enzyme
Catalyzes the polymerization
of deoxyribonucleotides into
a DNA strand.
Uses a magnesium ion for
catalytic activity.
DNA polymerases have
highly-conserved structure,
which means that their
overall catalytic subunits
vary, on a whole, very little
from species to species.

Function
DNA polymerase can add free nucleotides to
only the 3 end of the newly-forming strand. This
results in elongation of the new strand in a 5'-3'
direction.
Error correction(proof reading and repair) is a
property of some, but not all, DNA polymerases.
This process corrects mistakes in newlysynthesized

Different DNA polymerases have

different functions in the cell

Eukaryotic DNA Polymerases