title: New protein reagents. N-(4-Chloromercuriphenyl)-4-chloro-3,5-dinitrobenzamide and its use as a probe of the quaternary structure of yeast alcohol dehydrogenase.
abstract: The new bifunctional reagent, N-(4-chloromercuriphenyl)-4-chloro-3,5-dinitrobenzamide (I) was used to investigate the quaternary structure of yeast alcohol dehydrogenase. The four essential - SH groups of the enzyme were substituted by the mercuriphenyl moiety of compound I in the course of the reaction of one mole of protein with four moles of the reagent (one molecule of compound I incorporated by yeast alcohol dehydrogenase monomer). In a second step only two of the four chlorodinitrophenyl fragments bound to the protein established intermonomeric cross-links with non-essential - NH2 groups. The resulting dimers could be re-dissociated with mercaptoethanol. This result suggests that the four protomers of the enzyme could be arranged as a dimer of dimers.
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism; D001549:Benzamides; D007202:Indicators and Reagents; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D009243:NAD; D010663:Phenylmercury Compounds; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001549:Benzamides; D007202:Indicators and Reagents; D046911:Macromolecular Substances; D010663:Phenylmercury Compounds; D009243:NAD; C015065:N-(4-chloromercuriphenyl)-4-chloro-3,5-dinitrobenzamide; D000429:Alcohol Oxidoreductases