Kinetics, Regulation and Mechanisms of Biochemical Reactions

Chock, P B.

Abstract

(1) Experiments designed to test the direct transfer mechanism for metabolites in the glycolytic pathway have revealed that the data used to support the proposed mechanism are questionable. Contrary to the report that aldolase forms a complex with alpha- glycerol-3-phosphate dehydrogenase and facilitates the dehydrogenase catalyzed reaction through direct transfer of aldolase-bound substrate, dihydroxyacetone phosphate, we found that aldolase inhibits the dehydrogenase-catalyzed reaction by removing substrate for alpha-glycerol-3-phosphate dehydrogenase. In addition, transient kinetic data show that the reported first-ordered transfer rate of NADH from alpha- glycerol-3-phosphate dehydrogenase to lactate dehydrogenase is a composite of two reactions, one due to free NADH binding to lactate dehydrogenase and the other due to complex formation between the two dehydrogenases. (2) Binding constants for Ca(II) and a number of divalent metal ions have been determined for a family of Ca(II) indicators designed to probe intracellular calcium concentration. (3) A theory has been developed to account for the effect of oscillating of fluctuating membrane potential on the function of membrane-bound proteins. The theory provides a reasonable mechanism for energy transduction. In addition, an oscillating electric pulse apparatus is being constructed to carry out experiments to test the validity of the theory. (4) Work is in progress to purify and characterize isopeptidase.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Heart, Lung, and Blood Institute (NHLBI)
• Type: Intramural Research (Z01)
• Project #: 1Z01HL000202-17
• Application #: 3942760
• Support Year: 17
• Fiscal Year: 1987

Institution

• Name: U.S. National Heart Lung and Blood Inst
• Country: United States

Publications

Seo, Jae Ho; Lim, Jung Chae; Lee, Duck-Yeon et al. (2009) Novel protective mechanism against irreversible hyperoxidation of peroxiredoxin: Nalpha-terminal acetylation of human peroxiredoxin II. J Biol Chem 284:13455-65

Tanaka, Mikiei; Chock, P Boon; Stadtman, Earl R (2007) Oxidized messenger RNA induces translation errors. Proc Natl Acad Sci U S A 104:66-71

Li, Tianwei; Santockyte, Rasa; Shen, Rong-Fong et al. (2006) A general approach for investigating enzymatic pathways and substrates for ubiquitin-like modifiers. Arch Biochem Biophys 453:70-4

Miyoshi, Noriyuki; Oubrahim, Hammou; Chock, P Boon et al. (2006) Age-dependent cell death and the role of ATP in hydrogen peroxide-induced apoptosis and necrosis. Proc Natl Acad Sci U S A 103:1727-31

Li, Tianwei; Santockyte, Rasa; Shen, Rong-Fong et al. (2006) Expression of SUMO-2/3 induced senescence through p53- and pRB-mediated pathways. J Biol Chem 281:36221-7

Shelton, Melissa D; Chock, P Boon; Mieyal, John J (2005) Glutaredoxin: role in reversible protein s-glutathionylation and regulation of redox signal transduction and protein translocation. Antioxid Redox Signal 7:348-66

Khan, Mohammed A S; Chock, P Boon; Stadtman, Earl R (2005) Knockout of caspase-like gene, YCA1, abrogates apoptosis and elevates oxidized proteins in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 102:17326-31

Chiueh, Chuang C; Andoh, Tsugunobu; Chock, P Boon (2005) Roles of thioredoxin in nitric oxide-dependent preconditioning-induced tolerance against MPTP neurotoxin. Toxicol Appl Pharmacol 207:96-102

Oubrahim, Hammou; Wang, Jun; Stadtman, Earl R et al. (2005) Molecular cloning and characterization of murine caspase-12 gene promoter. Proc Natl Acad Sci U S A 102:2322-7

Tekle, Ephrem; Oubrahim, Hammou; Dzekunov, Sergey M et al. (2005) Selective field effects on intracellular vacuoles and vesicle membranes with nanosecond electric pulses. Biophys J 89:274-84