1.
Isozymes
Isozymes are different variants of the same enzyme having identical functions but different
amino acid sequence and kinetic parameters (e.g. different KM values), or are regulated
differently. In many cases, isozymes are encoded by homologous genes that have diverged
over time.
Even though isozymes are present in the same individual and catalyze the same reaction,
these reactions may take place under different cellular conditions or occur in different tissues
or cells.
For instance, acidity (pH), activation energy, temperature, and salinity may vary in different
tissues. An enzyme in the stomach would need to be more acid resistant than an enzyme that
catalyzes the same reaction in the vascular system.
A specific example of an isozyme is the enzyme Creatine kinase (CK), which has three forms:
CK-BB, CK-MB, and CK-MM.
CK-MB is found predominantly in cardiac tissue,
CK-BB in the brain, and
CK-MM in the skeletal tissue.
Elevations in CK-MB might indicate heart disease, such as a heart attack. When run through
gel electrophoresis, CK-BB is the most mobile while CK-MM is the least mobile.
Another example of an isozyme is glucokinase, a variant of hexokinase which is not inhibited
by glucose 6-phosphate. Its different regulatory features and lower affinity for glucose
(compared to other hexokinases), allow it to serve different functions in cells of specific
organs, such as
control of insulin release by the beta cells of the pancreas, or
initiation of glycogen synthesis by liver cells.
Both these processes must only occur when glucose is abundant.
2. Abzymes
An abzyme (from antibody and enzyme), also called catmab (from catalytic monoclonal
antibody) and most often called catalytic antibody or sometimes catab, is a monoclonal
antibody with catalytic activity.
Abzymes that are produced in vivo have binding specificity for specific substrates, but do
not have catalytic groups or display only weak, modest catalytic activity and have not
proved to be of any practical use. They are, however, subjects of considerable academic
interest. Studying them has yielded important insights into reaction mechanisms, enzyme
structure and function, catalysis, and the immune system itself.
Some abzymes have been engineered to use metal ions and other cofactors to improve
their catalytic activity.
Enzymes function by lowering the activation energy of the transition state of a chemical
reaction, thereby enabling the formation of an otherwise less-favorable molecular
intermediate between the reactant(s) and the product(s). If an antibody is developed to bind
to a molecule that is structurally and electronically similar to the transition state of a given
chemical reaction, the developed antibody will bind to, and stabilize, the transition state, just
like a natural enzyme, lowering the activation energy of the reaction, and thus catalyzing the
reaction. By raising an antibody to bind to a stable transition-state analog, a new and unique
type of enzyme is produced.
Potential HIV treatment:
In 2008, researchers at University Of Texas Medical School at Houston announced that they
have engineered an abzyme that degrades the superantigenic region of the gp120 CD4
binding site. This is the part of the HIV virus outer coating and is the attachment point to T
lymphocytes, the key cell in cell-mediated immunity. Once infected by HIV, patients produce
antibodies to the more changeable parts of the viral coat. The antibodies are ineffective
because of the virus' ability to change their coats rapidly. Because this protein gp120 is
necessary for HIV to attach, it does not change across different strains and is a point of
vulnerability across the entire range of the HIV variant population.
The abzyme does more than bind to the site: it catalytically destroys the site, rendering the
virus inert, and then can attack other HIV viruses. A single abzyme molecule can destroy
thousands of HIV viruses.
3. Ribozymes
Ribozymes (ribonucleic acid enzymes) are RNA molecules that have the ability to catalyze
specific biochemical reactions, including RNA splicing in gene expression, similar to the action
of protein enzymes.
The 1982 discovery of ribozymes demonstrated that RNA can be both genetic material
(like DNA) and a biological catalyst (like protein enzymes).
The most common activities of natural or in vitro evolved ribozymes are;
1. the cleavage (or ligation) of RNA and DNA and
2. peptide bond formation.
For example, within the ribosome, ribozymes function as part of the large subunit ribosomal
RNA to link amino acids during protein synthesis.
They also participate in a variety of RNA processing reactions, including RNA splicing, viral
replication, and transfer RNA biosynthesis.
Examples of ribozymes include the hammerhead ribozyme, the VS ribozyme, leadzyme, and
the hairpin ribozyme.
Applications of Ribozymes
Ribozymes have been proposed and developed for the treatment of disease through gene
therapy. One major challenge of using RNA-based enzymes as a therapeutic is the short
half-life of the catalytic RNA molecules in the body. To combat this, the 2’ position on the
ribose is modified to improve RNA stability. One area of ribozyme gene therapy has been
the inhibition of RNA-based viruses.
A type of synthetic ribozyme directed against HIV RNA called gene shears has been
developed and has entered clinical testing for HIV infection.
Similarly, ribozymes have been designed to target the hepatitis C virus RNA, SARS
coronavirus (SARS-CoV), Adenovirus and influenza A and B virus RNA. The ribozyme is
able to cleave the conserved regions of the virus's genome, which has been shown to
reduce the virus in mammalian cell culture. Despite these efforts by researchers, these
projects have remained in the preclinical stage.
