MORE ON ENZYMES

JULIUS P. MARIO, RMT, MS

ENZYME REGULATION 
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Control  Proenzymes  Allosterism  Protein Modification  Isoenzymes

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Formation

of the product inhibits an earlier reaction in a sequence the reaction, A  

In

E1

E2

E3

B

C

D

D may inhibit E1 activity by competitive or noncompetitive inhibition  When D is low, all three reactions proceed rapidly  When D is high, E1 becomes inhibited

thus. angiotensin  Trypsinogen .PROENZYMES  Inactive form of an enzyme  Also known as zymogen  Activated by cleavage of excess polypeptide chain  Cleavage of peptide promotes structural changes. functional  Examples are = cleavage of the 6 amino acids from the N-terminal by enteropeptidase converts it Nto active trypsin  Angiotensinogen = cleavage of the last 2 amino acids from the C-terminal by ACE converts it to a Cvasoactive octapeptide.

PROENZYMES  Chymotrypsinogen = cleavage of the 245 residues long polypeptide between R-15 and RI-16 from the N-terminal by trypsin converts it Nto active -chymotrypsin but the fully active form is -chymotrypsin .

PROENZYMES  Some enzymes have proenzymes because they are highly destructive when produced directly in their active forms these leak or exist in excess. inhibitory proteins bind them to render them inactive  Should .

ALLOSTERISM  If a substance binds noncovalently and reversibly to a site other than the active site.  It may inhibit the enzyme (Negative modulation)  It may stimulate enzyme action (Positive modulation) .

affect behavior is an allosteric effector  Allosteric effectors are substrates.ALLOSTERISM means other. activators. and inhibitors  Allo . steric means shape  Possible conformational changes affect the behavior of proteins  Due to multiple forms of the quaternary structure of some allosteric enzymes  A substance that modifies 4o structure and thus.

ALLOSTERIC PROTEINS  Proteins in which subtle changes at one site affects the structure and function of another site to cooperative effects as sigmoidal curve on enzyme  Due  Depicted kinetics  Examples are Aspartate transcarbamoylase (ATCase) & Hemoglobin .

dependent on the [S] with a maximal rate not dependent on [S] anymore as a hyperbolic curve on MichaelisMichaelis-Menten kinetics are chymotrypsin & myoglobin  Depicted  Examples .NONALLOSTERIC PROTEINS  At first.

protein. HETEROTROPIC Effects  Homotropic effects are allosteric interactions that occur when several identical molecules are bound to a protein.HOMOTROPIC vs. protein.  Heterotropic effects are allosteric interactions that occur when different substances are bound to a protein. .

typically by the addition of a functional group covalently bound to the apoenzyme example is the activation/inhibition of phosphorylation  Glycogen  Best-known Best- phosphorylase is active when it is phosphorylated at its serine or tyrosine residue  Pyruvate kinase from the liver is inactive when it is phosphorylated .PROTEIN MODIFICATION  Modification is usually a change in the primary structure.

ISOENZYMES  Enzymes that perform the same function but have different combinations of subunits. LD3 (H2M2). Km. and origins  Act on the same substrate  Have  LDH has 4 subunits  LD1 (H (H4). thus have different 4o structures different electrophoretic mobilities. LD2 (H3M). LD4 (H (H (HM3) & LD5 (M4) (M .

phenylalanyl.  Pepsin . leucyl or valyl isoleucyl. H & Q = same with chymotrypsin and others  Thermolysin = at isoleucyl. tryptophanyl or tyrosyl as well  Trypsin as L.CARBOXYPEPTIDASES  Cleave peptides and proteins at the carboxyl terminal of a particular amino acid in the chain = at lysyl or arginyl  Chymotrypsin = at phenylalanyl.

Aspartyl protease = a pair of aspartate side chains. a meat tenderizer) (papain.PROTEASE FAMILY   All members have similar chemical form Serine Proteases = cleaves at seryl residues chymotrypsin. sometimes on different subunits participate in the reaction (pepsin and HIV (pepsin protease) protease)   . trypsin and elastin) Cysteine protease = cleaves at cysteinyl residue (papain. elastin) (chymotrypsin.

ABZYMES  Antibodies elicited by antigenic proteins  Designer enzymes which can catalyze a wide variety of reactions  Usually are transition-state analogs which transitionwhen introduced into the body becomes immunogenic  N -(5 -phosphopyridoxyl)-L-lysine is phosphopyridoxyl)the transition-state analog for the reaction transitionbetween an amino acid and pyridoxal-5 pyridoxalphosphate .

glycolipid mannosyl transferases. 5 NT. hydroxymethylglutaryl-CoA reductase. cyt P-450 reductase. some 30 hydroxymethylglutarylenzymes for steroid synthesis and 20 enzymes of lipid synthesis  Luminal side = cyt P-450. ATPase. NADPHNADPH-ferrihemoprotein reductase.CELLULAR LEVEL  SER enzymes by the cytochrome P-450 P-  Detoxification enzymes  Lipid synthesis & degradation by side = cyt b5. cyt b5 reductase. PPglucoseglucose-6-phosphatase and -glucuronidase  Cytosolic .

5 -NT. NADPH:cyt c reductase. UDP-galactose-NUDP-galactoseacetylglucosamine. and many glycosylation enzymes . flippases for phospholipid synthesis  GOLGI BODY  Enzymes for posttranslational modifications of proteins synthesized on membranes as well as recycling of membrane material.-Dacetylglucosaminegalactosyltransferase. NADH:cyt c oxidoreductase. RER  Transport enzymes for glycosylation of newly formed proteins.

 LYSOSOMES  Hydrolytic enzymes such as proteinases. and sulfatases  MICROBODIES  Mainly oxidative enzymes  VACUOLES  V-type H+-translocating ATPase . lipases. glycosidases. phosphatases. nucleases.

BRUSH BORDER ENZYMES  Disaccharidases found on the microvilli of the small intestine  Once lost. can be re-synthesized via reenzyme induction commonly acted upon are maltose. and lactose  Substrates . sucrose.

Some pathogenic cocci are capable of producing hemolysins which interfere with oxygen transport in an organism  . The ammonia produced counters the harmful effects of the acid.ENZYMES AS VIRULENCE FACTORS  Helicobacter pylori survives the harsh gastric milieu by the urease on its cell wall.

Rifampin)  Binds .EFFECTS ON ENZYMES  Bactericidal antibiotics for both Gram positive and Gram negative organisms act via inhibition of cell wall synthesizing protein synthase (e. Carbapenems) to the subunit of RNA polymerase to inhibit transcription of mostly Gram+ bacteria & Mycobacteria (e.g.g.

cavernosum. the enzyme that degrades the signalling molecule cyclic guanosine monophosphate. This signalling molecule triggers smooth muscle relaxation and allows blood flow into the corpus cavernosum. it makes this signal last for a longer period of time. An example of a medicinal enzyme inhibitor is sildenafil (Viagra). a common treatment for male erectile dysfunction. monophosphate. which causes an erection. This compound is a potent inhibitor of cGMP specific phosphodiesterase type 5.   . Since the drug decreases the activity of the enzyme that halts the signal.

. Folic acid is the oxidised form of the substrate of dihydrofolate reductase. drug methotrexate to folic acid. Methotrexate blocks the action of dihydrofolate reductase and thereby halts thymidine biosynthesis. an enzyme that is potently inhibited by methotrexate. This block of nucleotide biosynthesis is selectively toxic to rapidly growing cells. therefore methotrexate is often chemotherapy. Another example of the structural similarity of some inhibitors to the substrates of the enzymes they target is seen in the figure comparing the acid. reductase. used in cancer chemotherapy.

This causes the cell wall to lose strength and the bacteria to burst. For example. needed for the survival of pathogens. Drugs also are used to inhibit enzymes pathogens. . bacteria are surrounded by a thick cell wall made of a net-like polymer netcalled peptidoglycan. Many antibiotics peptidoglycan. such as penicillin and vancomycin inhibit the enzymes (the transpeptidase from the bacteria Streptomyces R61) that produce and then cross-link the strands of this crosspolymer together.

 Humans  Selective toxicity is also produced in antibiotics by exploiting differences in the structure of the ribosomes in bacteria. do not make peptidoglycan. or how they make fatty acids . therefore inhibitors of this process are selectively toxic to bacteria. Drug design is facilitated when an enzyme that is essential to the pathogen's survival is absent or very different in humans.

acetate and choline. and chlorpyrifos irreversibly inhibit acetylcholinesterase. most. dopamine. an enzyme found in animals from insects to humans. including serotonin. edrophonium. parathion. in the treatment of myasthenia gravis and in anaesthesia. The carbamate pesticides are also examples of reversible AChE inhibitors. norepinephrine. are used neostigmine. AcCHE. and neostigmine. malathion.  Reversible competitive inhibitors. physostigmine.  The organophosphate insecticides such as malathion. and norepinephrine. parathion. It is essential to nerve cell function through its mechanism of breaking down the neurotransmitter acetylcholine into its constituents.  . such as inhibitors. choline. physostigmine. edrophonium. This is somewhat unique among neurotransmitters as serotonin. dopamine. are absorbed from the synaptic cleft rather than cleaved.

ENZYME DEFICIENCIES .

MUCOPOLYSACCHARIDOSIS MPS Type IH I H/S IS II III Eponym Deficient Enzyme aa-iduronidase a-iduronidase aaa-iduronidase Iduronidase sulfatase Heparan sulfatase N-acetylglucosaminidase Acetyl CoA glucosamine acetyltransferase N-acetylglucosamine-6-sulfatase acetylglucosamineGalactosamineGalactosamine-6-sulfatase b-galactosidase Hurler Hurler-Scheie HurlerScheie Hunter Sanfilippo A Sanfilippo B Sanfilippo C IV V VI VII IX Sanfilippo D Morquio A Morquio B Nonexistent MaroteauxMaroteaux-Lamy N-acetylhexosamine-4-sulfatase acetylhexosamineSly b-glucuronidase Hyaluronidase Hyaluronidase Deficiency .

lactic acidosis.All tissues debrancher enzyme (Cori-Forbes)Hepatomgaly. fasting hypoglycermia IV (Andersen). myoglobinuria . hyperlipidemia.4-glucosidase (Pompe)1.4Cardiomegaly. death in infancy V (McArdle)(McArdle)- Muscle phosphorylase Pain and stiffness after exertion.Liver and kidney Glucose-6-phosphatase Gierke) GlucoseHepatomegaly.Enzyme Deficient -Clinical Features I (von Gierke) .All tissues alpha 1. muscle weakness. severe fasting hypoglycemia II (Pompe). death in infancy and adults III (Cori-Forbes).GLYCOGEN STORAGE DISEASES Type . muscle weakness.All tissues brancher enzyme Portal cirrhosis.

VI (Hers) Liver phosphorylase Hepatomegaly. decerebration. high urinary catecholamines. mild fasting hypoglycemia Pain and stiffness on exertion Spasticity. death in infancy VII (Tarui) VIII Muscle and liver phosphofructokinase Brain and liver adenylate kinase IX X Hepatomegaly. occasional fasting hypoglycemia Liver and muscle cAMPcAMPHepatomegaly dependent kinase only Liver phosphorylase kinase .

LIPIDOSES a group of inherited disorders characterized by the accumulation of lipids in tissues especially the brain due to deficiency in a particular sphingolipid catabolic enzyme .

NiemannNiemann-Pick disease  Deficiency in sphingomyelinase and accumulation of sphingomyelin Gaucher s disease  Deficiency in F-D-glucosidase and accumulation of glucocerebroside .

Krabbe s disease  Deficiency in F-D-galactosidase and accumulation of galactocerebsides Fabry s disease  Deficiency in E-D-galactosidase and accumulation of ceramide trihexoside .

TayTay-Sach s disease  Deficiency in F-D-hexaminidase A and accumulation of ganglioside GM2 Metachromatic Leukodystrophy  Deficiency in sulfatide sulfatase and accumulation of Fsulfogalactocerebroside .

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