A KINETIC STUDY OF THE LIPOAMIDE DEHYDROGENASE-NADH-DYE REACTION.
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)
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 1965
- Accession Number
- AD0615169
Entities
People
- David N. Kramer
- George G. Guilbault
- Paul Goldberg
Organizations
- Edgewood Chemical Biological Center