Cofactors
-a non-protein chemical compound that is required for
the enzyme's biological activity.
-a co-catalyst essential for enzyme activity.
-Thus, a cofactors can be considered as a "helper
molecules" that assist enzymes in their action.
-Apoenzyme - enzyme lacking cofactor (inactive)
Holoenzyme - enzyme with cofactors (active)
-an apoenzyme is the inactive protein without a cofactor
Cofactors
classification
Organic molecules- also
Inorganic molecules such as
called coenzyme such as
metal ions like Zn+2, Mg+2 etc
NADH, TPP,
They influence the substrate binding and
participate in catalytic reactions or play the role
of an electron carrier.
Some enzymes require both a coenzyme and one or more metal
ions for activity.
The cofactors are again subdivided into
-prosthetic groups
-cosubstrates.
When a coenzyme or metal ion is very tightly or
even covalently bound to the enzyme protein, it is
called a prosthetic group.
-The prosthetic group is bound firmly to the
enzyme.
-It can not be removed by, e. g. dialysis,
-During enzyme catalysis it remains attached to the
enzyme molecule.
Example: Biotin, FADH, FMN, pyridoxal phosphate
Loosely attached to apoenzymes, seperated easily by
dialysis they are often called co-substrate or
secondary substrate.
Example: NAD+ , ATP
Metal ions that are tightly bound to enzyme and
participate directly in catalysis (Fe, Zn, Cu, Co).
While those that are not tightly bound are required
for activation or stimulation of the enzyme or/and
often participate in substrate binding.
Example:
Mg2+, is required by many ATP requiring enzymes
such as 6 CH O H 6 CH O PO 2
2 2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H 2+ OH
Mg
OH OH OH OH
3 2 3 2
H OH H ex ok in ase H OH
glu co se glu co se -6 -p h osp h ate
Example of metal ions that
are prosthetic group
Metalloenzymes contain
firmly bound metal ions
at the enzyme active
sites (examples: iron,
Example of metalloenzyme:
zinc, copper, cobalt). carbonic anhydrase contains
zinc
Vitamin Insufficiency Generally Result in malfunction of
enzymes
Metalloenzymes
Metalloenzymes are enzyme proteins containing metal ions (metal
cofactors), which are directly bound to the protein or to enzyme-
bound nonprotein components (prosthetic groups).
-About one-third of all enzymes known so far are metalloenzymes.
-- Zinc is required for the activity of > 300 enzymes, covering all six
classes of enzymes.
-These metalloenzymes are capable of catalyzing various classes of
important reactions in biosynthesis and metabolism .
-These metal ions mainly act as Lewis acids or as redox-active sites.
-metal ions are usually coordinated
by nitrogen, oxygen or sulfur centers belonging to amino
acid residues of the protein.
The hammerhead ribozyme is a metalloenzyme; Mg+2 ions are required for
activity. Are autocatalytic RNA molecule that cleaves single stranded RNA.
-carbonic anhydrase contains zinc ions at its active site.
-Pyruvate kinase:
-4 metal binding sites
-Mg2+ and K+
Alpha Amylase:
-Calcium ion stabilizes the structure.
Catalases-
protect the cell from oxidative damage by
excess hydrogen peroxide produced during O2 metabolism,
contains Fe ion at its catalytic centre.
-superoxide dismutase:
contains zinc and cu ions
--arginase in urea cycle
- Mn2+
Nitrogenase:
-molybdenum
-Fe-S centre.
Numerous binuclear metalloenzymes are involved in
digestion and metabolism of protein and peptides.
example: aminopeptidase a member of the zinc
α,β-hydrolases superfamily contains two zinc ions at
its active site.
Bovine leucine aminopeptidase (BLAP) is
another classical binuclear metalloprotease containing
two zinc ions in the active-site.
Certain enzymes have been employed in both laboratory reactions
and industrial-scale reactions to synthesize valuable organic
compounds.
In one notable example, nitrile hydratase, which includes a Co(III)
ion in the reactive site, has been used for industrial production of
acrylamide.
Isoenzyme
Different enzymes That Catalyze the Same Reaction
may occur in the same species, in the same
tissue, or even in the same cell.
The different forms of the enzyme generally differ in
kinetic (V and K ) or regulatory properties,
max M
cofactor they use
their subcellular distribution (soluble or membrane-
bound).
Isozymes may have similar, but not identical, amino acid
sequences,
in many cases they clearly share a common evolutionary
origin.
coded by different genes.
Example
Lactate dehydrogenase (LDH) exists as at least five different
isozymes separable by electrophoresis.
LDH1-HHHH -Heart and erythrocyte
LDH2 -HHHM -Heart and erythrocyte
LDH3- HHMM -Brain and kidney
LDH4 -HMMM -Skeletal muscle and liver
LDH5 -MMMM -Skeletal muscle and liver
Hexokinase
Hekinase I- myocytes
Hexikinase II-myocytes
Hexokinase III
Hexokinase IV
Why are different form of enzymes exist?
Different metabolic patterns in different [Link] glycogen phosphorylase,
the isozymes in skeletal muscle and liver have different regulatory properties,
reflecting the different roles of glycogen breakdown in these two tissues.
Different locations and metabolic roles for isozymes in the same cell. The
isocitrate dehydrogenase isozymes of the cytosol and the mitochondrion are
an example.
Different stages of development in embryonic or fetal tissues and in adult
tissues. For example, the fetal liver has a characteristic isozyme distribution of
LDH, which changes as the organ develops into its adult form. Some enzymes
of glucose catabolism in malignant (cancer) cells occur as their fetal, not adult,
isozymes.
Different responses of isozymes to allosteric modulators. This difference is
useful in fine-tuning metabolic rates. Hexokinase IV (glucokinase) of liver and
the hexokinase isozymes of other tissues differ in their sensitivity to inhibition
by glucose 6-phosphate.
