Abstract
The purpose of this study was to investigate the effects exerted by thiol-modifying reagents on themitochondrial sulfonylurea receptor. The thiol-oxidizing agents (timerosal and 5, 5'-dithio-bis(2-nitrobenzoic acid)) were found to produce a large inhibition (70% to 80%) of specific binding of [(3)H]glibenclamide to the beef heart mitochondrial membrane. Similar effects were observed with membrane permeable (N-ethylmaleimide) and non-permeable (mersalyl) thiol modifying agents. Glibenclamide binding was also decreased by oxidizing agents (hydrogen peroxide) but not by reducing agents (reduced gluthatione, dithiothreitol and the 2,3-dihydroxy-1,4-dithiolbutane). The results suggest that intact thiol groups, facing the mitochondrial matrix, are essential for glibenclamide binding to the mitochondrial sulfonylurea receptor.
Copyright 1999 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ATP-Binding Cassette Transporters*
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Adenosine Diphosphate / pharmacology
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Adenosine Triphosphate / pharmacology
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Animals
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Binding Sites / drug effects
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Cattle
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Dithionitrobenzoic Acid / pharmacology
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Dose-Response Relationship, Drug
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Ethylmaleimide / metabolism
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Ethylmaleimide / pharmacology
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Glyburide / metabolism
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Hydrogen Peroxide / pharmacology
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Intracellular Membranes / drug effects
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Intracellular Membranes / metabolism
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Kinetics
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Mersalyl / pharmacology
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Mitochondria, Heart / drug effects*
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Mitochondria, Heart / metabolism
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Oxidants / pharmacology
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Permeability
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Potassium Channels / metabolism*
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Potassium Channels, Inwardly Rectifying*
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Receptors, Drug / metabolism*
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Reducing Agents / pharmacology
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Sulfhydryl Compounds / metabolism
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Sulfhydryl Reagents / pharmacology*
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Sulfonylurea Receptors
Substances
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ATP-Binding Cassette Transporters
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Oxidants
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Potassium Channels
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Potassium Channels, Inwardly Rectifying
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Receptors, Drug
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Reducing Agents
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Sulfhydryl Compounds
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Sulfhydryl Reagents
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Sulfonylurea Receptors
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Mersalyl
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Adenosine Diphosphate
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Adenosine Triphosphate
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Dithionitrobenzoic Acid
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Hydrogen Peroxide
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Ethylmaleimide
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Glyburide