This proposal concerns the time-dependent changes in chemical structure and in protein conformation during the transient catalyzed reactions involving the enzymes, histidinol-PO4 amino transferase (HAT) and cation translocating ATPases, and two enzymes of the glycolytic pathway, GPDH and PGK. One proposed study is to use mutant enyzme species of the HAT to effect different distributions of reaction intermediates during catalysis and, consequently, upon chemical identification of intermediates to define the catalyzed reaction pathway. A chromophoric 32P-ATP analog will be utilized in conjunction with intrinsic cation signals to investigate the coupled mechanism of energy coupled vectorial transport. The glycolytic pathway will be examined in regard to the coupled reaction sequence catalyzed by GPDH and PGK. Since interactions between the two enzymes have been demonstrated, the relevance to metabolism and the role of effectors on this interaction will be investigated.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM010451-21
Application #
3268113
Study Section
Biochemistry Study Section (BIO)
Project Start
1976-05-01
Project End
1986-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
21
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Oregon
Department
Type
Graduate Schools
DUNS #
948117312
City
Eugene
State
OR
Country
United States
Zip Code
97403
Srivastava, D K; Bernhard, S A (1987) Biophysical chemistry of metabolic reaction sequences in concentrated enzyme solution and in the cell. Annu Rev Biophys Biophys Chem 16:175-204
Srivastava, D K; Bernhard, S A (1986) Metabolite transfer via enzyme-enzyme complexes. Science 234:1081-6
Srivastava, D K; Bernhard, S A (1986) Enzyme-enzyme interactions and the regulation of metabolic reaction pathways. Curr Top Cell Regul 28:1-68
Srivastava, D K; Bernhard, S A (1985) Mechanism of transfer of reduced nicotinamide adenine dinucleotide among dehydrogenases. Biochemistry 24:623-8
Srivastava, D K; Bernhard, S A; Langridge, R et al. (1985) Molecular basis for the transfer of nicotinamide adenine dinucleotide among dehydrogenases. Biochemistry 24:629-35
Senear, D F; Betts, G; Bernhard, S A (1985) Multiple ion-dependent and substrate-dependent Na+/K+-ATPase conformational states. Transient and steady-state kinetic studies. Biochemistry 24:6789-98