Binding of Rat Brain Hexokinase to Recombinant Yeast Mitochondria: Effect of Environmental Factors and the Source of Porin |
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Authors: | Claude Aflalo Heftsiba Azoulay |
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Institution: | (1) Department of Life Sciences, The Ben Gurion University of the Negev, P.O. Box 653, Beer Sheva, 84105, Israel |
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Abstract: | Heterologous binding of rat brain hexokinase to wild type, porinless, and recombinant yeast mitochondria expressing human porin was assessed, partially characterized, and compared to that in the homologous system (rat liver mitochondria). With porin-containing yeast mitochondria it is shown that (i) a significant, saturatable association occurs; (ii) its extent and apparent affinity, correlated with the origin of porin, are enhanced in the presence of dextran; (iii) the binding requires Mg ions and apparently follows a complex cooperative mechanism. This heterologous association does not seem to differ fundamentally from that in the homologous system and represents a good basis for molecular studies in yeast. With porinless yeast mitochondria, binding occurs at much lower affinity, but to many more sites per mitochondrion. The results indicating a major but not exclusive role for porin in the binding are discussed in terms of (i) the mode and mechanism of binding, and (ii) the suitability of the rat hexokinase–yeast mitochondria couple for the study of heterogeneous catalysis in reconstituted cellular model systems. |
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Keywords: | Mitochondria rat brain hexokinase porin VDAC yeast cooperative binding |
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