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β-sulfopyruvate: Chemical and enzymatic syntheses and enzymatic assay

descriptionPublicationkeyboard_double_arrow_right Article 01 Jul 1986Publisher:Elsevier BVJournal:Analytical Biochemistry, volume 156, pages 154-160 (issn: 0003-2697,

Authors: Catherine L. Weinstein; Owen W. Griffith;

doi: 10.1016/0003-2697(86)90167-3

pmid: 3740406

β-sulfopyruvate: Chemical and enzymatic syntheses and enzymatic assay

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Abstract

beta-Sulfopyruvic acid (2-carboxy-2-oxoethanesulfonic acid) is prepared in greater than 90% yield by reaction of bromopyruvic acid with sodium sulfite. beta-[35S]Sulfopyruvate is prepared by transamination between [35S]cysteinesulfonate (cysteate) and alpha-ketoglutarate using mitochondrial aspartate aminotransferase isolated from rat liver. Following either chemical or enzymatic synthesis, the crude reaction product is conveniently purified by chromatography on Dowex 1; beta-sulfopyruvate is isolated as the stable, water-soluble dilithium salt. beta-Sulfopyruvate is shown to be an alternative substrate of mitochondrial malate dehydrogenase; in the presence of 0.25 mM NADH, beta-sulfopyruvate is reduced with an apparent Km of 6.3 mM and a Vmax equal to about 40% of that observed with oxaloacetate. This finding forms the basis of a convenient spectrophotometric assay of beta-sulfopyruvate.

Related Organizations

New York University
United States
Cornell University
United States

Keywords

Chromatography, Mitochondria, Liver, Sulfur Radioisotopes, Rats, Kinetics, Malate Dehydrogenase, Isotope Labeling, Animals, Spectrophotometry, Ultraviolet, Aspartate Aminotransferases, Pyruvates, Oxidation-Reduction, Cysteic Acid

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