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.
| 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 |
