Isozymes or isoenzymes

• Many enzymes are present in more than one molecular form in the
same species, in the same tissue or even in the same cell.
• Such forms of the enzyme which catalyse the same reaction but
exhibit different kinetic properties are called isoenzymes.
• They can be separated by electrophoresis since they have different
amino acid compositions.
• Eg. Lactate dehydrogenase(LDH), creatine phosphokinase(CPK),
alkaline phosphatase etc.
 Lactate dehydrogenase

• LDH exists in five different forms in various tissues, LDH1, LDH2,

LDH3,LDH4 and LDH5.
• Two major types of LDH found are heart LDH and Muscle LDH.
• Heart LDH is found in the heart tissue and active at low levels of
pyruvate.
• It has four identical subunits called H subunit.
• Muscle LDH is present in skeletal muscles and is active in high
concentrations of pyruvate.
• It has also four subunits called M subunits.
• The H and M subunits possess the same molecular weight but
different amino acid composition. LDH isoenzymes differ at the level
of quaternary structure.
• LDH1 – H4
• LDH2- H3 M
• LDH3- H2M2
• LDH4- HM3
• LDH5- M4
• Creatinine phosphokinase(CPK) exists in three forms
• Each enzyme is a dimer and each subunit may be of M (muscle) type
or B(brain) type.
• They are CPK1(BB), CPK2(MB) and CPK3(MM).
• CPK1 is found in brain,CPK2 in the myocardium and CPk3 in skeletal
muscle.
• CPK2 is present in very small amount in the normal serum.
• Following myocardial infarction,its level rises within 4 hours and
reaches to maximum within one day.
• Alkaline phosphatase exists in three forms
 zymogen

• Some enzymes are found in the form of inactive precursors.

• Such forms are called proenzymes or zymogens.they become active
after conversion to original form.
• The prefix or ‘pro’ suffix ‘-ogen’ is added to enzymes name to denote
its zymogen form.
• Eg. Pepsinogen,trypsinogen,prothrombin
• Activation of trypsinogen: trypsinogen is secreted by pancreas.
• An intestinal enzyme enterokinase converts trypsinogen to trypsin
• Trypsinogen enterokinase trypsin + hexapeptide
 Abzymes (catalytic antibodies)
• Abzymes are antibodies that catalyse specific chemical reactions ie, they
can perform dual role, as an antibody and an enzyme.
• Abzymes cut peptide bonds at specific amino acid residues, such as
restriction enzymes cut DNA at specific sites.
• Eg. Monoclonal antibodies acting as enzyme
• The binding of an antibody to its antigen can be viewed as an
enzyme-substrate reaction.
• In both cases the binding is a non-covalent, weak interaction, but with high
specificity and affinity.
• The binding also lowers the activation energy.
• But normally the antibody does not alter the antigen, as the enzyme does
the subsrate.
• Exploring the possibility of using an antibody to alter an antigen as
enzymes do is the latest in biotechnological research.
• eg.to raise monoclonal antibodies murine spleen cells have been
immunized with a hapten-carrier complex in which the hapten
structurally resembles the transition state of an ester undergoing
hydrolysis.
• Such anti hapten Mabs can accelerate the hydrolysis of an ester
substrate 1000 fold, which means the mabs act like the enzyme
catalysing the substrates hydrolysis.
• such catalytic antibodies are infact produced is the strongest
evidence in support of the transition theory of enzyme action.
 Ribozymes

• Enzymes made up of RNA are called ribozymes.

• They are catalytic RNA molecules with sequence specific cleavage
activity.
• They show kinetics typical of enzymes.
• Thomas Cech and Sidney Altman discovered the ribozymes for which
they were awarded Nobel prize in 1989.
• Ribozymes catalyse two reactions
• 1. cleavage of RNA and
• 2. cleavage of DNA
• They are able to cut and splice themselves into a form that can catalyse the
cleavage of other RNA/DNA molecules.
• The catalytic activity of ribozymes arises due to their three dimensional
structures which is able to generate in them the subsrate-specific binding
site.
• This is similar to enzymes in which the substrate binding site is produced by
their three-dimensional structure.
• All biological catalysts must possess the ability to bind to their specific
substrates.
• Eg. Ribonuclease p, peptidyl transferase present in ribosomes
• Applications- in human, animal and plant disease control.