Central University of South Bihar
Panchanpur, Gaya, India
E-Learning Resources
Department of Biotechnology
NB: These materials are
taken/borrowed/modified/compiled from various sources
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teaching purpose in a public university, and for serving
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MSBTN2003C04
Unit 4
Structure and function of enzymes
By
Dr Rakesh Kumar
Department of Biotechnology
(Lecture 6)
Note: These materials are only for classroom teaching purpose at
13/4/2020
CUSB Gaya.
Isoenzymes
• Isozymes (also known as isoenzymes or more generally as Multiple
forms of enzymes) are enzymes that differ in amino acid sequence but
catalyze the same chemical reaction.
• Isozymes were first described by R. L. Hunter and Clement Markert (1957)
who defined them as different variants of the same enzyme having
identical functions and present in the same individual.
• These enzymes usually display different kinetic parameters (i.e. different
KM values), or different regulatory properties
• They have similar catalytic activity, but are different biochemically or
immunologically.
• Two or more polypeptide chains
• Different polypeptide chains are products of different genes
• Different forms may be differentiated from each other based on certain
physical properties
– electrophoretic mobility,
– differences in absorption properties
Origin of isozymes
• Isozymes are usually the result of gene duplication, but can
also arise from polyploidisation or nucleic acid hybridization.
Over evolutionary time, if the function of the new variant
remains identical to the original, then it is likely that one or the
other will be lost as mutation accumulate, resulting in
a pseudogene. However, if the mutations do not immediately
prevent the enzyme from functioning, but instead modify
either its function, or its pattern of gene expression, then the
two variants may both be favoured by natural selection and
become specialised to different functions. For example, they
may be expressed at different stages of development or in
different tissues.
• Isozymes represent enzymes from different genes that
process or catalyse the same reaction
(1) enzyme variants that are the product of different
genes and thus represent different loci (described
as isozymes) and
(2) enzymes that are the product of different alleles of
the same gene (described as allozymes).
Example of an isozyme
Lactate Dehydrogenase (LD)
•Catalyzes interconversion of lactic and pyruvic acids
•It is a hydrogen-transfer enzyme
•NAD is used as coenzyme
•High activities in heart, liver, muscle, kidney, and RBC
•Lesser amounts: Lung, smooth muscle and brain.
•LDH occurs as a tetramer of 2 different subunits H & M: product of 2 diff.
gene)
Contd…
• The enzyme, as found in many species, is a tetramer of molecular
weight 140000. however, although each subunit has a molecular weight
of about 35 000, two types, of different amino acid composition, are
found within each species: that are the M form which predominates in
skeletal muscle and other largely anaerobic tissues; and the H form, the
predominates sub unit in the heart. The two types of subunit are
produced by separate gene. Each monomer is catalytically inactive, but
it can combine with others of the same or different type to produce the
active tetrameric enzyme. All combinations of H and M subunits are
equally possible, so five isoenzyme of LDH can exist: H 4, H3M, H2M2,
HM3, and M4 (termed LDH1 – LDH5).
• 5 isoenzymes composed of a cardiac (H) and muscle ( M ) component
– LD - 1 ( HHHH ) Cardiac , RBCs
– LD - 2 ( HHHM ) Cardiac , RBCs
– LD - 3 ( HHMM ) Brain and Kidney, Lungs
– LD - 4 ( HMMM ) Hepatic
– LD - 5 ( MMMM ) Skeletal muscle
Isoenzyme Composition Composition Present in Elevated in
name
LDH1 ( H 4) HHHH Myocardium, myocardial
RBC infarction
LDH2 (H3M1) HHHM Myocardium,
RBC
LDH3 (H2M2) HHMM Brain and
Kidney, Lungs
LDH4 (H1M3) HMMM Hepatic
LDH5 (M4) MMMM Skeletal Skeletal muscle
muscler and liver
diseases
Alkaline phosphatase
• Alkaline phosphatase (ALP, ALKP) is a hydrolase enzyme responsible
for removing phosphate groups from many types of molecules,
including nucleotides, proteins, and alkaloids.
• The process of removing the phosphate group is called
dephosphorylation.
• As the name suggests, alkaline phosphatases are most effective in
an alkaline environment. It is sometimes used synonymously as basic
phosphatase.
• In humans, alkaline phosphatase is present in all tissues throughout the
entire body, but is particularly concentrated in liver, bile duct, kidney, bone,
and the placenta. Humans and most other mammals contain the following
alkaline phosphatase isozymes:
– ALPI – Intestinal
– ALPL – tissue non-specific (liver/bone/kidney)
– ALPP – placental
CREATINE KINASE (CK)
• Creatine kinase is associated with ATP
regeneration in muscle and nervous tissue
Creatine + ATP phosphocreatine + ADP
(Phosphocreatine – serves as energy reserve during muscle contraction)
Creatine kinase is a dimer made of 2 monomers
occurs in the tissues
Skeletal muscle contains M subunit, Brain contains B
subunits
Three different isoenzymes are formed
Isoenzyme
Composition Present in Elevated in
name
CK-1 BB Brain CNS diseases
Acute
CK-2 MB Heart myocardial
infarction
CK-3 MM Muscle
Significance of isozyme
• In a particular tissue a particular isoenzyme of this enzyme may be
present. If that tissue is damaged ,the analysis of the blood level of
isoenzymes of Lactic Dehydrogenase, can determine which tissue is
damaged. This is because the isoenzyme corresponding to that tissue will
be raised in the blood.
