The Role of an Essential Histidine Residue of Yeast Alcohol Dehydrogenase

1975 ◽  
Vol 52 (3) ◽  
pp. 595-603 ◽  
Author(s):  
Christopher J. DICKENSON ◽  
F. Mark DICKINSON
Keyword(s):  
Alcohol Dehydrogenase ◽  
Histidine Residue ◽  
Yeast Alcohol Dehydrogenase

scholarly journals A study of the ionic properties of the essential histidine residue of yeast alcohol dehydrogenase in complexes of the enzyme with its coenzymes and substrates

1977 ◽  
Vol 161 (1) ◽  
pp. 73-82 ◽  
Author(s):  
C J Dickenson ◽  
F M Dickinson
Keyword(s):  
Alcohol Dehydrogenase ◽  
Initial Rate ◽  
Dissociation Constants ◽  
Ternary Complexes ◽  
Histidine Residue ◽  
Mechanism Of Formation ◽  
Binary And Ternary Complexes ◽  
Yeast Alcohol Dehydrogenase ◽  
Ph Range

1. Initial-rate studies of the reduction of acetaldehyde by NADH, catalysed by yeast alcohol dehydrogenase, were performed at pH 4.9 and 9.9, in various buffers, at 25 degrees C. The results are discussed in terms of the mechanism previously proposed for the pH range 5.9-8.9 [Dickenson & Dickinson (1975) Biochem. J. 147, 303-311]. 2. Acetaldehyde forms a u.v.-absorbing complex with glycine. This was shown not to affect the results of kinetic experiments under the conditions used in this and earlier work. 3. The variation with pH of the dissociation constant for the enzyme-NADH complex, calculated from the initial-rate data, indicates that the enzyme possesses a group with pK7.1 in the free enzyme and pK8.7 in the complex. 4. The pH-dependences of the second-order rate constants for inactivation of the enzyme by diethyl pyrocarbonate were determined for the free enzymes (pK7.1), the enzyme-NAD+ complex (pK approx. 7.1) and the enzyme-NADH complex (pK approx. 8.4). The essential histidine residue may therefore be the group involved in formation and dissociation of the enzyme-NADH complex. 5. Estimates of the rate constant for reaction of acetaldehyde with the enzyme-NADH complex indicate that acetaldehyde may combine only when the essential histidine residue is protonated. The dissociation constants for butan-1-ol and propan-2-ol, calculated on the basis of earlier kinetic data, are, however, independent of pH. 6. The results obtained are discussed in relation to the role of the essential histidine residue in the mechanism of formation of binary and ternary complexes of the enzyme with its coenzymes and substrates.

Role of histidine in molecule of pea alcohol dehydrogenase

1982 ◽  
Vol 47 (5) ◽  
pp. 1408-1413 ◽  
Author(s):  
Noemi Čeřovská ◽  
Jana Barthová ◽  
Sylva Leblová
Keyword(s):  
Alcohol Dehydrogenase ◽  
Enzymatic Activity ◽  
Inactivation Rate ◽  
Histidine Residue ◽  
Michaelis Constant ◽  
Diethyl Pyrocarbonate ◽  
Inactivation Rate Constant ◽  
Pea Seedlings ◽  
Absorbance Difference

Alcohol dehydrogenase (E.C.1.1.1.1) from germinating pea seedlings was modified by treatment with diethyl pyrocarbonate. The inactivation rate is proportional to the molar concentration of the modifying agent; the inactivation was almost complete in fifty minutes at a diethyl pyrocarbonate concentration of 5 . 10-6 mol/l. The histidine content calculated from the absorbance difference at 240 nm was 3.43 residues per molecule of native and 4.75 residues per molecule of demetalized enzyme. A correlation of the absorbance difference at 240 nm with a 100% loss of enzymatic activity shows that 1.22 histidine residue is essential for the activity of alcohol dehydrogenase. The dependence of the inactivation rate constant on the pH of the medium indicates that the treatment of pea alcohol dehydrogenase with diethyl pyrocarbonate results in the modification of one group only with a pK of 7.1, well corresponding to the imidazole group of histidine. The enzyme is partially protected against inactivation by NADH at a concentration close to the Michaelis constant for NADH. The treatment of the ethoxyformylated enzyme with hydroxylamine followed by dialysis restored the activity of pea alcohol dehydrogenase by 88%.

Role of metal ions to specific binding of yeast alcohol dehydrogenase by free and immobilized dyes

1983 ◽  
Vol 79 ◽  
pp. 157-158
Author(s):  
S.S. Flaksaite ◽  
O.F. Sudzhiuvene ◽  
J.-H. J. Pesliakas ◽  
A.A. Glemzha
Keyword(s):  
Alcohol Dehydrogenase ◽  
Metal Ions ◽  
Specific Binding ◽  
Yeast Alcohol Dehydrogenase

Inhibitory and destabilizing role of putrescine on yeast alcohol dehydrogenase

2011 ◽  
Vol 44 (13) ◽  
pp. S86-S87
Author(s):  
Darbandsar Mazandarani Samira ◽  
Moosavi-Nejad Zahra
Keyword(s):  
Alcohol Dehydrogenase ◽  
Yeast Alcohol Dehydrogenase

The Role of Histidine Residues of Yeast Alcohol Dehydrogenase

1973 ◽  
Vol 1 (6) ◽  
pp. 1270-1272 ◽  
Author(s):  
CHRISTOPHER J. DICKENSON ◽  
F. MARK DICKINSON
Keyword(s):  
Alcohol Dehydrogenase ◽  
Histidine Residues ◽  
Yeast Alcohol Dehydrogenase

scholarly journals Role of the essential thiol groups of yeast alcohol dehydrogenase

1972 ◽  
Vol 126 (1) ◽  
pp. 133-138 ◽  
Author(s):  
F. M. Dickinson
Keyword(s):  
Alcohol Dehydrogenase ◽  
Dissociation Constant ◽  
Binding Capacity ◽  
Ternary Complexes ◽  
Native Enzyme ◽  
Thiol Groups ◽  
Yeast Alcohol Dehydrogenase

1. Yeast alcohol dehydrogenase inactivated by reaction with iodoacetamide retains 85% of the original NADH-binding capacity as measured under conditions of saturating coenzyme concentration. 2. The dissociation constant of the enzyme–NADH complex is unaffected by inactivation of the enzyme with iodoacetamide, and the affinity of the enzyme for NAD+ and pyridine-3-aldehyde–adenine dinucleotide (PAAD+) appears to be similarly unaffected. 3. Enzyme inactivated with iodoacetamide has lost the ability to form normal ternary complexes of the type enzyme–NADH–acetamide and enzyme–PAAD+–hydroxylamine that are characteristic of the native enzyme.

scholarly journals Evidence for a histidine and a cysteine residue in the substrate-binding site of yeast alcohol dehydrogenase

1975 ◽  
Vol 145 (3) ◽  
pp. 581-590 ◽  
Author(s):  
V Leskovac ◽  
D Pavkov-Peričin
Keyword(s):  
Alcohol Dehydrogenase ◽  
Binding Site ◽  
Ternary Complex ◽  
Substrate Binding ◽  
Cysteine Residue ◽  
Histidine Residue ◽  
Thiol Groups ◽  
Substrate Binding Site ◽  
Yeast Alcohol Dehydrogenase ◽  
Diethyl Pyrocarbonate

1. Yeast alcohol dehydrogenase (EC 1.1.1.1) is inhibited by stoicheiometric concentrations of diethyl pyrocarbonate. The inhibition is due to the acylation of a single histidine residue/monomer (mol.wt. 36000). 2. Alcohol dehydrogenase is also inhibited by stoicheiometric amounts of 5,5′-dithiobis-(2-nitrobenzoate), owing to the modification of a single cysteine residue/monomer. 3. Native alcohol dehydrogenase binds two molecules of reduced coenzyme/molecule of enzyme (mol.wt. 144000). 4. Modification of a single histidine residue/monomer by treatment with diethyl pyrocarbonate prevents the binding of acetamide in the ternary complex, enzyme-NADH-acetamede, but does not prevent the binding of NADH to the enzyme. 5. Modification of a single cysteine residue/monomer does not prevent the binding of acetamide to the ternary complex. After the modification of two thiol groups/monomer by treatment with 5,5′-dithiobis-(2-nitrobenzoate), the capacity of enzyme to bind coenzyme in the ternary complex was virtually abolished. 6. From the results presented in this paper we conclude that at least one histidine and one cysteine residue are closely associated in the substrate-binding site of alcohol dehydrogenase.

Sign in / Sign up

Export Citation Format

Share Document