Calorimetric demonstration of the potential of molecular crowding to emulate the effect of an allosteric activator on pyruvate kinase kinetics

Lonhienne, T. G. A. and Winzor, D. J. (2002) Calorimetric demonstration of the potential of molecular crowding to emulate the effect of an allosteric activator on pyruvate kinase kinetics. Biochemistry, 41 22: 6897-6901.


Author Lonhienne, T. G. A.
Winzor, D. J.
Title Calorimetric demonstration of the potential of molecular crowding to emulate the effect of an allosteric activator on pyruvate kinase kinetics
Journal name Biochemistry   Check publisher's open access policy
ISSN 0006-2960
Publication date 2002
Sub-type Article (original research)
DOI 10.1021/bi020064h
Volume 41
Issue 22
Start page 6897
End page 6901
Total pages 5
Place of publication Washington, USA
Publisher American Chemical Society
Collection year 2002
Language eng
Subject C1
270100 Biochemistry and Cell Biology
780105 Biological sciences
Abstract A method based on isothermal calorimetry is described for the direct kinetic assay of pyruvate kinase. In agreement with earlier findings based on the standard coupled assay system for this enzyme in the presence of a fixed ADP concentration, the essentially rectangular hyperbolic dependence of initial velocity upon phosphoenolpyruvate concentration is rendered sigmoidal by the allosteric inhibitor phenylalanine. This effect of phenylalanine can be countered by including a high concentration of a space- filling osmolyte such as proline in the reaction mixtures. This investigation thus affords a dramatic example that illustrates the need to consider potential consequences of thermodynamic nonideality on the kinetics of enzyme reactions in crowded molecular environments such as the cell cytoplasm.
Keyword Biochemistry & Molecular Biology
Mediated Conformational-changes
Thermodynamic Nonideality
Isomerization Equilibrium
Enzyme-kinetics
Inert Solutes
Volume
Preexistence
Hydrolysis
Cosolutes
Linkages
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Biomedical Sciences Publications
 
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