Document Type

Article

Publication Date

9-1-1993

Publication Title

Archives of Biochemistry and Biophysics

Abstract

Initial velocity studies obtained with alternative dinucleotide substrates for the 6-phosphogluconate dehydrogenase reaction suggest that the 2′-phosphate is critical for the optimum productive binding of the dinucleotide substrate. Initial velocity patterns obtained by varying 6-phosphogluconate at different fixed levels of NAD are nearly parallel with apparent competitive substrate inhibition by 6-phosphogluconate at pH 7 and below but intersect to the left of the ordinate at pH 8 and above. Dead-end inhibition studies indicate that the mechanism is random at all pH values. Data are interpreted in terms of a random mechanism with marked antagonism in the binding of NAD and 6-phosphogluconate at low pH. Deuterium isotope effects on V and V/K for either substrate are equal at pH 8, indicating that the kinetic mechanism is rapid equilibrium random. A decrease in the pH and the subsequent protonation of the active site general base or some other enzyme residue with a similar pK apparently results in the ineffective binding of NAD. The latter suggests either a link between the protonation state of this group and the conformation of the dinucleotide binding site or an interaction between the two.

Comments

This work was supported by grants to P.F.C. from NIH (GM 36799) and the Robert A. Welch Foundation (B-1031) and to A.J.B. from Sigma Xi (GIAR Award 9203).

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

DOI

10.1006/abbi.1993.1460

Version

Postprint

Volume

305

Issue

2

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