beta-Sulfopyruvate: chemical and enzymatic syntheses and enzymatic assay.
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.
D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D002845:Chromatography / Q000379:methods; D003544:Cysteic Acid / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D007700:Kinetics; D008291:Malate Dehydrogenase / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D011773:Pyruvates / Q000032:analysis / Q000096:biosynthesis / Q000138:chemical synthesis*; D051381:Rats; D013056:Spectrophotometry, Ultraviolet; D013462:Sulfur Radioisotopes
D011773:Pyruvates; D013462:Sulfur Radioisotopes; C049788:beta-sulfopyruvic acid; D003544:Cysteic Acid; D008291:Malate Dehydrogenase; D001219:Aspartate Aminotransferases