ALLOSTERIC REGULATION &

COVALENT MODIFICATION

Bhaskar Ganguly
Ph.D. , M.V.Sc., B.V.Sc. & A.H.

Allosteric Regulation
Modulator binds to the allosteric site of an enzyme to alter
its kinetic characteristics; may be either stimulatory or
inhibitory. Stimulator is often the substrate itself and such
enzymes are called homotropic.
When the modulator and substrate differ, the enzyme is called
heterotropic. Some enzymes have more than one
modulators.
The allosteric site is specific for its modulator. Enzymes with
several modulators generally have different specific binding
sites for each. By contrast, in homotropic enzymes, the active
site and allosteric site are the same.

a. The V vs. [S] curve for allosteric enzyme is typically sigmoid as

compared for hyperbolic curves for non-allosteric enzymes. b. Effect

Allosteric Regulation
Two general models for the interconversion of inactive and
active forms of allosteric enzymes have been proposed:
Simple sequential model and Concerted/ Symmetry model.
In the concerted model, the binding of substrate to one of
the subunits increases the probability that both switch
from the T to the R form. Thus, symmetry is conserved in
the concerted model but not in the sequential model.

Covalent Modification
Protein modifications include di-sulfide linkages,
acetylation,
hydroxylation,
phosphorylation,
methylation, glycosylation and even the addition of
nucleotides.
Acetylation is the most common form, affecting 80 % of all
proteins.
Hydroxylation of Pro is common in collagen.
Actively transcribed chromatin is associated with the
acetylation of lysine residues in the N-terminal regions of the
core histones, whereas the condensation of chromosomes at
mitosis is accompanied by the phosphorylation of histone H1.
Glycoproteins are commonly involved in cell recognition.

Covalent modifications of nucleic acids have specific roles

in the cell. In DNA, the methylation of adenine and
cytosine bases is common.
The methylation of CpG is associated with transcriptionally
inactive regions of chromatin.
Hemi-methylation of the DNA after replication allows the
daughter strand to be distinguished from the parental strand.

Protein Phosphorylation
The phosphorylation and dephosphorylation of amino acid
side chains by protein kinases and phosphatases provide
reversible on/off regulation of numerous proteins.
Serine,
Threonine
and
Tyrosine
are
commonly
phosphorylated residues.
Nearly 3 percent of all yeast proteins are protein kinases or
phosphatases.
Mouse has 540 different kinases. The
human genome encodes at least 500 protein kinases and
100 different phosphatases.

Protein Kinases
Serine/ Threonine Kinases: Phosphorylate serine/
threonine residues; e.g. PKA, PKB, MAP Kinase (ERK),
etc.
Protein Kinase A (PKA):

Virtually all the diverse effects of cAMP are mediated through

protein kinase A (PKA), also called cAMP-dependent protein
kinase.
Although PKA acts on different substrates in different types of
cells, it always phosphorylates a serine or threonine residue
that occurs within the same sequence motif: X-Arg-(Arg/Lys)X-(Ser/Thr)-$, where X denotes any amino acid and $ denotes
a hydrophobic amino acid.

Protein Kinase B (PKB):

Protein kinase B (PKB) becomes partially activated by binding

to PI 3-phosphates; full activation requires phosphorylation by
another kinase (PDK1).
Activated protein kinase B promotes survival of many cells by
directly inactivating several pro-apoptotic proteins and downregulating expression of others.

Protein Kinases
Receptor Tyrosine Kinases: Auto-phosphorylate
internal tyrosine residues. Downstream signaling
involves Ras and serine/ threonine kinases, ultimately
leading to activation of MAP kinase and transcription
factors.

Thank you