title: Redox specificity of 2-hydroxyacid-coupled NAD(+)/NADH dehydrogenases: a study exploiting "reactive" arginine as a reporter of protein electrostatics.
abstract: With "reactive" arginine as a kinetic reporter, 2-hydroxyacid dehydrogenases are assessed in basis of their specialization as NAD(+)-reducing or NADH-oxidizing enzymes. Specifically, M4 and H4 lactate dehydrogenases (LDHs) and cytoplasmic and mitochondrial malate dehydrogenases (MDHs) are compared to assess if their coenzyme specificity may involve electrostatics of cationic or neutral nicotinamide structure as the basis. The enzymes from diverse eukaryote and prokaryote sources thus are assessed in "reactivity" of functionally-critical arginine as a function of salt concentration and pH. Electrostatic calculations were performed on "reactive" arginines and found good correspondence with experiment. The reductive and oxidative LDHs and MDHs are assessed in their count over ionizable residues and in placement details of the residues in their structures as proteins. The variants found to be high or low in ΔpKa of "reactive" arginine are found to be also strong or weak cations that preferentially oxidize NADH (neutral nicotinamide structure) or reduce NAD(+) (cationic nicotinamide structure). The ionized groups of protein structure may thus be important to redox specificity of the enzyme on basis of electrostatic preference for the oxidized (cationic nicotinamide) or reduced (neutral nicotinamide) coenzyme. Detailed comparisons of isozymes establish that the residues contributing in their redox specificity are scrambled in structure of the reductive enzyme.
authors.affiliation: Department of Chemistry, Indian Institute of Technology Bombay, Mumbai, India.
authors.forename: Pooja
authors.initials: P
authors.lastname: Gupta
authors.affiliation: Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Mumbai, India ; Department of Biosciences, Jamia Millia Islamia, New Delhi, India.
authors.forename: Mohamad Aman
authors.initials: MA
authors.lastname: Jairajpuri
authors.affiliation: Department of Chemistry, Indian Institute of Technology Bombay, Mumbai, India.
authors.forename: Susheel
authors.initials: S
authors.lastname: Durani
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D011817:Rabbits; D055672:Static Electricity; D013552:Swine
chemical_list: D007527:Isoenzymes; D009243:NAD; D001120:Arginine; D000429:Alcohol Oxidoreductases; C083779:2-hydroxyacid dehydrogenase; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase