A comprehensive presentation on Enzymology :Types of Enzyme inhibition & Therapeutic uses for MBBS ,BDS, B Pharm & Biotechnology students to facilitate self- study.
6. Irreversible inhibition
• Irreversible inhibitor : destroys a functional group on enzyme
necessary for catalytic activity
• eg Di Isopropyl Fluorophosphate(DFP ) inhibits Acetyl choline
esterase
• Iodoacetamide inhibits OH group of serine ,SH group of Cysteine
,Imidazole group of Histidine
7. DFP – inhibits Serine Proteases such as Acetyl Choline
Esterase , Trypsin ,Chymotrypsin ,Elastase
BAL –BRITISH ANTI LEVISITE- Antidote for HEAVY METALS
CYANIDE –Cytochrome Oxidase
FLUORIDE –inhibits Enolase ( removes Mg ⁺⁺/Mn⁺⁺ from the
active site OF enzymes
Irreversible inhibition of enzymes
11. Irreversible inhibition of enzymes with SH group -by Iodoacetamide
enzymes with SH group + Iodoacetamide
reversible inhibition of enzymes with Chemically modified enzyme ( inactive )
14. Allosteric Regulation
a) Allosteric inhibitor-binds to an
allosteric site on enzyme &
induces conformation change preventing
Substrate to bind to an active site on enzyme
.Product formation inhibited .
16. Allosteric & covalent modulation
Activation of many
enzymes involve
Phosphorylation of
enzyme molecule by
Protein Kinases.
Deactivation of many
enzymes is facilitated
by dephosphorylization
of enzyme by
Phosphoprotein
Phosphatases
17. Reversible inhibition of enzymes by Competitive inhibitors
Competitive inhibitors
1. Competitive inhibitors competes with substrate for binding to
active site but once bound substrate cannot be transformed into
product by enzymes.
2. Inhibition by Competitive inhibitors can be reversed by simply
increasing concentration of substrate
3. Competitive inhibitors resembles the normal substrate in 3D
structure
4. E + I = EI
18. NO PRODUCT FORMATION
Comparison of
competitive and
non-competitive
inhibition
Competitive
inhibitors
resembles the
normal substrate in
3D structure
&non-Competitive
inhibitors don’t
resembles the
normal substrate in
3D structure
19. COMPETITIVE INHIBITION ---Competitive inhibitors
1 .competes with
substrate for binding to
active site but once
bound substrate cannot
be transformed into
product by enzymes.
2.Inhibition by
Competitive inhibitors
can be reversed by
simply increasing
concentration of
substrate
20. Changes in Reaction Rate in presence of Competitive & Non –competitive inhibitor
Inhibition by Competitive inhibitors
can be reversed by simply increasing
concentration of substrate& that by
non -Competitive inhibitors
cann’t be reversed by simply
increasing substrate concentration.
23. Structure of Succinic acid ,Malonic acid & oxaloacetic acid with two carboxylic acid groups
(COOH )& can bind to an active site on succinate dehydrogenase –Competitive inhibitors
28. Non competitive Inhibition
Characteristics of Non competitive Inhibition
1. Reversible but not reversed by substrate
2. Inhibitor binds at site other than substrate binding site
3. It binds reversibly to both free enzyme & ES complex to form
inactive complex EI & ESI
E + I ↔ EI
ES + I ↔ ESI
4. Inhibitors alter the conformation of E molecule so that reversible
activation occurs
5. They are naturally occurring metabolic intermediates .
29. Non competitive inhibition
Threonine lsoleucine ( Threonine Dehydratase inhibited by isoleucine-final
product of cascade /pathway ) -feed back inhibition
30. Comparison between competitive & Non competitive inhibition
Criteria competitive Non competitive
Active on Active site May or may not be active at binding
site
Structure of inhibitors Substrate analogs Unrelated molecules
Inhibition reversible Generally irreversible
Excess of substrate Inhibition relieved No effect
Km Increased in presence of
inhibitor
Unchanged in presence of inhibitor
Vmax Unchanged decreased
Significance Therapeutic application Toxicological application
31. Comparison between competitive & Non competitive inhibition
Competition inhibition –Vmax unchanged & Km INCREASED
Non competitive inhibition –Vmax decreased & Km not altered
32. Comparison between competitive & non competitive inhibition
Catalytic site
Regulatory
site
Isoleucine
Binds to
regulatory site on
Threonine
Dehydratase &
functions as non-
competitive
inhibitor
34. Uncompetitive inhibition of enzymes
Uncompetitive inhibition- inhibitors binds to ES Complex
Vmax & Km decreased eg Alkaline Phosphatase by phenylalanine
35. Suicide Inhibition
Properties of Suicide Inhibition
1. Irreversible inhibition
2. More effective inhibitor
3. synthesized with the help of enzyme
4. Inhibition of xanthine oxidase by Allopurinol (treatment of GOUT )
5. Alloxanthine synthesized by xanthine oxidase using Allopurinol is more
potent inhibitor of enzyme than Allopurinol
6. Arachidonic acid Prostaglandin ( cyclo oxygenase )is inhibited by Aspirin
.(anti inflammatory )
7. 5 Fluoro-Uracil 5 Fluoro Deoxy Uridylate more potent inhibitor of
Thymidylate synthase to inhibit nucleotide synthesis and is being used in cancer
treatment