title: Chemical modification of arginine residues of rat liver S-adenosylhomocysteinase.
abstract: Rat liver S-adenosylhomocysteinase (EC 3.3.1.1) is inactivated by phenylglyoxal following pseudo-first order kinetics. The dependence of the apparent first order rate constant for inactivation on the phenylglyoxal concentration shows that the inactivation is second order in reagent. This fact together with the reversibility of inactivation upon removal of excess reagent and the lack of reaction at residues other than arginine as revealed by amino acid analysis and incorporation of phenylglyoxal into the protein indicate that the inactivation is due to the modification of arginine residue. The substrate adenosine largely but not completely protects the enzyme against inactivation. Although the modification of two arginine residues/subunit is required for complete inactivation, the relationship between loss of enzyme activity and the number of arginine residues modified, and the comparison of the numbers of phenylglyoxal incorporated into the enzyme in the presence and absence of adenosine indicate that one residue which reacts very rapidly with the reagent compared with the other is critical for activity. Although the phenylglyoxal treatment does not result in alteration of the molecular size of the enzyme or dissociation of the bound NAD+, the intrinsic protein fluorescence is largely lost upon modification. The equilibrium binding study shows that the modified enzyme apparently fails to bind adenosine.
mesh_terms: D043383:Adenosylhomocysteinase; D000447:Aldehydes / Q000494:pharmacology*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001120:Arginine; D001665:Binding Sites; D006867:Hydrolases / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D010658:Phenylglyoxal / Q000494:pharmacology*; D011485:Protein Binding; D051381:Rats; D013050:Spectrometry, Fluorescence
chemical_list: D000447:Aldehydes; D000596:Amino Acids; D001120:Arginine; D006867:Hydrolases; D043383:Adenosylhomocysteinase; D010658:Phenylglyoxal