Accession Number : AD0615169

Title :   A KINETIC STUDY OF THE LIPOAMIDE DEHYDROGENASE-NADH-DYE REACTION.

Descriptive Note : Rept. for Jan-Jun 64,

Corporate Author : CHEMICAL RESEARCH AND DEVELOPMENT LABS EDGEWOOD ARSENAL MD

Personal Author(s) : Guilbault,George G. ; Goldberg,Paul ; kramer,David N.

Report Date : APR 1965

Pagination or Media Count : 20

Abstract : Because of the inhibition of the enzyme lipoamide dehydrogenase, diaphorase, by sulfhydryl binding compounds, interest developed in the mechanism by which this enzyme acts on various substrates. The application of fluorometric and spectrophotometric techniques to study the kinetics of the diaphorase-NADH-dye reaction is illustrated. Differences are observed in the kinetic relationships depending on the synthetic substrate: resazurin or dichloroindophenol. For resazurin, initial reaction velocities are independent of NADH concentration, but in the instance of the indophenols, a dependence on NADH was found. These results have been interpreted on the basis of a particular enzymatic reaction scheme, in which the substrate is reduced by a reduced form of the enzyme, rather than by an enzyme-NADH complex. (Author)

Descriptors :   (*OXIDOREDUCTASES, REACTION KINETICS), (*NUCLEOTIDES, REACTION KINETICS), (*DYES, REACTION KINETICS), CHEMICAL INDICATORS, FLUORESCENCE, COLORIMETRIC ANALYSIS, NITROGEN HETEROCYCLIC COMPOUNDS, RESORCINOL, PHENOLS

Distribution Statement : APPROVED FOR PUBLIC RELEASE