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ABZYMES, ISOENZYMES,
RIBOZYMES
B. C. Muthubharathi
2016501012
Abzymes
 Antibodies/Enzymes
 Catmab (catalytic Monoclonal antibody)
 Abzymes = Antibody +Enzymes
Similarity and Differe...
 1986 - 1st monoclonal catalytic antibodies (abzymes) against a
chemically stable analog of the transition state of a rea...
Sources
 Do not occur naturally in human
 Found in normal human ( Anti Vasoactive intestinal peptide
autoantibody which ...
Protoabzymes
 Proteolytic activity
 Found in AD patients – Asthuma, Multiple sclerosis, Thyroiditis,
Myocarditis
DNA abz...
Production
 Antibody molecules are produced by the immune system to
bind and neutralize foreign substances called antigen...
Antibody Antigen Neutralize antigen
Foreign protein / Hapten
TSA act as a hapten at TS and elicit antibody production, Ant...
Reactions
of
abzymes
Trans
esterificati
on
Photo
cleavage
Oxidatio
nDecarboxylat
ion
Cyclizatio
n
Hydrolysis
of enol
ether...
Hydrolysis of hydroxyester
 Hydroxyester forms cyclic intermediate during hydrolysis
Hydroxyester Cyclic intermediate  l...
Hydrolysis of ester
 Ester –Tetrahedral intermediate
P- nitrophenylacetate Tetrahedral p- nitrophenyl phosphate  Acetic
...
Pyridoxal 5’-P dependet aminotransferase reaction
D alanine +pyridoxal 5’-p Pyruvate + pyridoxamine 5’ –p
Catalyze
Ab rais...
Biosynthesis of heme ( Metallation)
 Introduction of ferrous ion into protoporphyrin
 Ferrochelatase
 Mechanism – Disto...
Activation of anticancer drug by
abzymes
Thymidylate synthase Building block of DNA
5- fluorodeoxyuridasine-5 phosphate
Ab...
ISOENZYMES
 Same reaction but differ in aminoacid sequences, kinetic properties
and electrophoretic mobility
 ONTOGENETI...
List of isoenzymes
Lactate dehydrogenase
Hexokinase
Creatin phosphokinase
Phosphorylase
Alkaline phosphatase
Cytochrome P4...
LACTATE DEHYDROGENASE
 Tetramer / oligomer (Different polypeptide chain)
 Four polypeptide subunit
 2 types(H- Heart, M...
 Glycolysis Pyruvate Lactate Blood
LDH
LDH5 (M4) work best in NAD+ - regenerating direction
Because, activate muscle tiss...
Hexokinase
 4 Isoenzymes
 I, II, III, IV
 I – Fastest mobility towards anode
 IV – Slowest mobility towards anode (als...
ANGIOTENSIN CONVERTING
ENZYME
 2 ISOENZYME
Somatic isoenzymes
Germinal isoenzymes
Somatic – Lung, Vascular endothelial ce...
Creatine phosphokinase
2 isoforms
B- Brain
M- Muscle
Catalyze phosphorylation of crreatine
Creatine + ATP creatine phospha...
RIBOZYMES
RIBONUCLEIC ACID
Not protein
Antisense RNAmolecules
8 natural ribozymes
1989 nobel prize, Thomas Czech
Catalytic...
 separate catalytic substrate-binding domains
 Hammerhead, Hairpin for Therapeutic purpose
 Hammerhead ribozymes are pr...
 In cell culture, ribozymes inhibit the expression of
(1) human cytomegalovirus transcriptional regulatory
proteins, resu...
Abzymes, ribozymes
Abzymes, ribozymes
Abzymes, ribozymes
Abzymes, ribozymes
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Abzymes, ribozymes

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this will be useful to understand about the new topics such as abzymes, ribozymes and also isoenzymes. You have to clear that ribozymes are not protein. because all enzymes are proteins but all proteins are not enzymes except ribozymes

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Abzymes, ribozymes

  1. 1. ABZYMES, ISOENZYMES, RIBOZYMES B. C. Muthubharathi 2016501012
  2. 2. Abzymes  Antibodies/Enzymes  Catmab (catalytic Monoclonal antibody)  Abzymes = Antibody +Enzymes Similarity and Differences  Enzymes - bind in high energy state  Antibody - binds the complementary structure in its ground state  ability to bind with compounds with great specificity and high affinity
  3. 3.  1986 - 1st monoclonal catalytic antibodies (abzymes) against a chemically stable analog of the transition state of a reaction were obtained  Peter Schultz and Richard Lerner  Having structural complementarity for the transition state of an enzyme catalyzed reaction  They bind strongly to the transition state with high association constant, enhancing the reaction rate  Abzymes reduce rotational entropy
  4. 4. Sources  Do not occur naturally in human  Found in normal human ( Anti Vasoactive intestinal peptide autoantibody which detect the level of vasoactive intestinal peptide) Natural abzymes in Patients with AD Blood of Asthuma – Reduce the rotational entropy Autoimmune diseased person (SLE)
  5. 5. Protoabzymes  Proteolytic activity  Found in AD patients – Asthuma, Multiple sclerosis, Thyroiditis, Myocarditis DNA abzymes DNA hydrolyzing activity Powerful regulator of apoptosis ( pathogenic role is not clear) Cytotoxicity mechanism in systemic autoimmune disease & Tumors
  6. 6. Production  Antibody molecules are produced by the immune system to bind and neutralize foreign substances called antigens  Foreign proteins of bacteria , viruses and some chemical molecules act as antigens  Transition state – State corresponding to highest potential energy along this reaction coordinates  Transition state analog- Molecules which are more stable than the transition state itself, but they mimic their 3D structure
  7. 7. Antibody Antigen Neutralize antigen Foreign protein / Hapten TSA act as a hapten at TS and elicit antibody production, Antibody isolated from serum used as abzymes Substrate Transition state Product Transition analog
  8. 8. Reactions of abzymes Trans esterificati on Photo cleavage Oxidatio nDecarboxylat ion Cyclizatio n Hydrolysis of enol ethers Reduction of diketone
  9. 9. Hydrolysis of hydroxyester  Hydroxyester forms cyclic intermediate during hydrolysis Hydroxyester Cyclic intermediate  lactone Phenol Increasing the reaction rate Anti cyclic intermediate antibody Cyclic phosphonate ester (Ag) ( Mimic cyclic intermediate)  Hydroxyester forms cyclic intermediate during hydrolysis Hydroxyester Cyclic intermediate  lactone Phenol Increasing the reaction rate Anti cyclic intermediate antibody Cyclic phosphonate ester (Ag) ( Mimic cyclic intermediate)
  10. 10. Hydrolysis of ester  Ester –Tetrahedral intermediate P- nitrophenylacetate Tetrahedral p- nitrophenyl phosphate  Acetic acid intermediate Binding (Catalysis) Antibodies against phosphate analog of ester phosphate analog of ester
  11. 11. Pyridoxal 5’-P dependet aminotransferase reaction D alanine +pyridoxal 5’-p Pyruvate + pyridoxamine 5’ –p Catalyze Ab raised N- (5’ phosphopyridoxyl)- lysine Coupled with carrier protein
  12. 12. Biosynthesis of heme ( Metallation)  Introduction of ferrous ion into protoporphyrin  Ferrochelatase  Mechanism – Distortion of pyrole ring by 36 to create bent transition state Protoporphyrin Bent transition state heme (product) N methyl mesoporphyrin Ab complementary (TSA) to TS
  13. 13. Activation of anticancer drug by abzymes Thymidylate synthase Building block of DNA 5- fluorodeoxyuridasine-5 phosphate Abzymes 5- fluorouracil (Anticancer drug)
  14. 14. ISOENZYMES  Same reaction but differ in aminoacid sequences, kinetic properties and electrophoretic mobility  ONTOGENETIC – Same species – Intraspecific variants  PHYLOGENETIC – Different species - Interspecific variants  HOMOLOGUES – Similar molecular structure &catalytic property Eg.Phospholipase D  ANALOGOUS - Different molecular structure but catalyzing same reaction Eg. Phenylalanine transaminase
  15. 15. List of isoenzymes Lactate dehydrogenase Hexokinase Creatin phosphokinase Phosphorylase Alkaline phosphatase Cytochrome P450 G - 6 – P dehydrogenase Glutamate dehydrogenase Differ in amino acid composition and kinetic behavior so, by using electrophoretic method, isoenzymes can be separated
  16. 16. LACTATE DEHYDROGENASE  Tetramer / oligomer (Different polypeptide chain)  Four polypeptide subunit  2 types(H- Heart, M- Muscle)  H4, H3M, H2M2, HM3, M4 (Catalyze the same reaction)  Pyruvate + NADH + H+ Lactate + NAD H4 LDH1 Heart Muscle H3M LDH2 RBC, Brain Muscle, Liver H2M2 LDH3 Brain, Leucocyte Heart, Liver HM3 LDH4 Leucocyte Heart, RBC,Brain Muscle, Liver M4 LDH5 Muscle, Liver Other tissues Increased level Decrerased levlel
  17. 17.  Glycolysis Pyruvate Lactate Blood LDH LDH5 (M4) work best in NAD+ - regenerating direction Because, activate muscle tissue become anaerobic HEART TISSUE Aerobic Lactate as fuel Lactate Pyruvate Citric acidCycle LDH1(H4) inhibited by excess pyruvate DIAGNOSTIC ENZYME ( increased level) Heart attack, Hemolytic anemia, Muscle damage, Muscular dystrophy, Stroke, Lung tissues, Death, hepatitis
  18. 18. Hexokinase  4 Isoenzymes  I, II, III, IV  I – Fastest mobility towards anode  IV – Slowest mobility towards anode (also called as Glucokinase)  I – Brain, Heart, Kidney  II – Skeletal muscle, Adipose tissue  III – Liver, Spleen  IV – Liver D- Glucose + ATP D – Glucose + ADP
  19. 19. ANGIOTENSIN CONVERTING ENZYME  2 ISOENZYME Somatic isoenzymes Germinal isoenzymes Somatic – Lung, Vascular endothelial cells, Renal epithelial cells, Testicular epithelial cells, Testicular epithelial cells Germinal – Sperm Regulate the blood pressure
  20. 20. Creatine phosphokinase 2 isoforms B- Brain M- Muscle Catalyze phosphorylation of crreatine Creatine + ATP creatine phosphate CPK I – BB - Brain, Lungs CPK II – MB – Heart CPK III – MM- Skeletal muscle
  21. 21. RIBOZYMES RIBONUCLEIC ACID Not protein Antisense RNAmolecules 8 natural ribozymes 1989 nobel prize, Thomas Czech Catalytic RNA molecules Ribonucleic acid enzymes/ RNA enzymes, RNAzymes RNA metaloenzymes ~40 to 50 nucleotides in length Sid Altman Tom Cech
  22. 22.  separate catalytic substrate-binding domains  Hammerhead, Hairpin for Therapeutic purpose  Hammerhead ribozymes are preferable Ability to more efficiently recognize, Bind to, and cleave a range of different mRNAs.  By altering the substrate-binding domain, a ribozyme can be engineered to specifically cleave any mRNA sequence  Inhibit the expression of a variety of viral genes and significantly inhibit the proliferation of numerous organisms  Mang Yu &Coworkers  Use ribozymes to provide WBC with resistance to HIV infection
  23. 23.  In cell culture, ribozymes inhibit the expression of (1) human cytomegalovirus transcriptional regulatory proteins, resulting in a 150-fold decrease in viral growth; (2) human herpes simplex virus type1 transcriptional activator, resulting in a reduction of around 1,000-fold in viral growth; and (3) A reovirus mRNA encoding a protein required for viral proliferation  Hammerhead – treat collagen-induced arthritis in mice

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