Modification of yeast pyruvate kinase by an active site-directed reagent, bromopyruvate.

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Modification of yeast pyruvate kinase by an active site-directed reagent, bromopyruvate.

The reaction of bromopyruvate with pyruvate kinase was demonstrated both by noting loss of catalytic activity with time and by incorporation of [14C]pyruvate. The first order rate constant for loss of catalytic activity when plotted as a function of bromopyruvate concentration fit a regular hyperbolic saturation function indicative of the formation of a complex with Ki = 15 mM prior to the inac...

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Allosteric properties of yeast pyruvate kinase studied by site-directed mutagenesis.

The enzyme from most sources is a tetramer of identical subunits each about 500 residues in length. The enzyme has an absolute requirement for monovalent and divalent cations (usually K+ and Mg2+) which act to coordinate and orientate the substrates prior to catalysis. A number of isoenzymes of pyruvate kinase are found to exist in vertebrate tissues. Their kinetic and regulatory properties ref...

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The active site of yeast phosphoglycerate kinase.

Eby, D. & Kirtley, M. E. (1971) Biochemistry 10,2677-2682 Fuller-Noel, J. K. & Schumaker, V. W. (1972) J. Mol. Biol. 68,523-532 Gennis, L. S. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 3928-3932 Harrigan, P. J. & Trenthami, D. R. (1971) Biochem. J . 124, 573-580 Hams, J. I. & Polgar, L. (1965) J. Mol. Biol. 14, 630-633 Harris, J. I. & Waters, M. (1976) Enzymes 3rd Ed. 23, 1-50 Kirschner, K. (1971...

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Yeast Pyruvate Kinase

The kinetic properties of purified yeast pyruvate kinase were investigated. The enzyme showed cooperative kinetics toward the essential activating monovalent cations K+ and NH4+, Mg2+, and phosphoenolpyruvate. Fructose 1,6diphosphate, which yielded homotropic cooperative kinetics and did not affect maximal velocity, transformed the sigmoidal kinetics of Kf and NH4+, Mgz+, and phosphoenolpyruvat...

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Asp295 Stabilizes the Active-Site Loop Structure of Pyruvate Dehydrogenase, Facilitating Phosphorylation of Ser292 by Pyruvate Dehydrogenase-Kinase

We have developed an in vitro system for detailed analysis of reversible phosphorylation of the plant mitochondrial pyruvate dehydrogenase complex, comprising recombinant Arabidopsis thalianaα2β2-heterotetrameric pyruvate dehydrogenase (E1) plus A. thaliana E1-kinase (AtPDK). Upon addition of MgATP, Ser292, which is located within the active-site loop structure of E1α, is phosphorylated. In add...

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ژورنال

عنوان ژورنال: Journal of Biological Chemistry

سال: 1979

ISSN: 0021-9258

DOI: 10.1016/s0021-9258(17)37727-x