Effect of pH on pyrazole binding to liver alcohol dehydrogenase. 1981

P Andersson, and J Kvassman, and A Lindström, and B Oldén, and G Pettersson

1. Kinetic and equilibrium data have been determined at different pH between 4 and 10 for binding of the inhibitor pyrazole to liver alcohol dehydrogenase and to the binary complexes formed between enzyme and NADH or NAD+. 2. Pyrazole binding to free enzyme requires the protonated form of an ionizing group with a pKa of 9.2, agreeing with the pKa value reported for the water molecule bound at the catalytic zinc ion of the enzyme subunit. The rate of association of the inhibitor to the enzyme . NAD+ complex exhibits a similar pKa-7.6-dependence attributable to ionization of zinc-bound water in the latter binary complex. These observations lend support to the idea that pyrazole combines to the catalytic zinc ion on complex formation with the enzyme, zinc-bound water most likely being displaced by the inhibitor. 3. The rate of dissociation of the inhibitor from the ternary enzyme . NAD+ . pyrazole complex is proportional to the hydrogen ion concentration over the examined pH range (4-8). This effect of pH, which is proposed to reflect ionization of the enzyme-bound inhibitor with a pKa value below 4 (indirectly estimated to 2.4), accounts for the exceptional stability of the ternary complex at neutral and alkaline pH. It is concluded that pyrazole, by analogy to water and alcohol ligands, undergoes a drastic pKa perturbation on binding to the catalytic zinc ion in the enzyme . NAD+ complex.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D009243 NAD A coenzyme composed of ribosylnicotinamide 5'-diphosphate coupled to adenosine 5'-phosphate by pyrophosphate linkage. It is found widely in nature and is involved in numerous enzymatic reactions in which it serves as an electron carrier by being alternately oxidized (NAD+) and reduced (NADH). (Dorland, 27th ed) Coenzyme I,DPN,Diphosphopyridine Nucleotide,Nadide,Nicotinamide-Adenine Dinucleotide,Dihydronicotinamide Adenine Dinucleotide,NADH,Adenine Dinucleotide, Dihydronicotinamide,Dinucleotide, Dihydronicotinamide Adenine,Dinucleotide, Nicotinamide-Adenine,Nicotinamide Adenine Dinucleotide,Nucleotide, Diphosphopyridine
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D011720 Pyrazoles Azoles of two nitrogens at the 1,2 positions, next to each other, in contrast with IMIDAZOLES in which they are at the 1,3 positions.
D006736 Horses Large, hoofed mammals of the family EQUIDAE. Horses are active day and night with most of the day spent seeking and consuming food. Feeding peaks occur in the early morning and late afternoon, and there are several daily periods of rest. Equus caballus,Equus przewalskii,Horse, Domestic,Domestic Horse,Domestic Horses,Horse,Horses, Domestic
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000429 Alcohol Oxidoreductases A subclass of enzymes which includes all dehydrogenases acting on primary and secondary alcohols as well as hemiacetals. They are further classified according to the acceptor which can be NAD+ or NADP+ (subclass 1.1.1), cytochrome (1.1.2), oxygen (1.1.3), quinone (1.1.5), or another acceptor (1.1.99). Carbonyl Reductase,Ketone Reductase,Carbonyl Reductases,Ketone Reductases,Oxidoreductases, Alcohol,Reductase, Carbonyl,Reductase, Ketone,Reductases, Carbonyl,Reductases, Ketone
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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