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..-(/sup 35/S)Sulfopyruvate is prepared by transamination between (/sup 35/)cysteinesulfonate (cysteate) and ..cap 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 K/sub m/ of 6.3 mM and a V/sub max/ equal to about 40% of that observed with oxaloacetate. This finding forms the basis of a convenient spectrophotometric assay of ..beta..-sulfopyruvate.
- Research Organization:
- Cornell Univ. Medical College, New York, NY (USA)
- OSTI ID:
- 6151721
- Journal Information:
- Anal. Biochem.; (United States), Vol. 156
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
OXIDOREDUCTASES
BIOCHEMICAL REACTION KINETICS
SUBSTRATES
AMINOTRANSFERASES
ION EXCHANGE CHROMATOGRAPHY
LABELLED COMPOUNDS
SPECTROPHOTOMETRY
SULFUR 35
TRACER TECHNIQUES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ENZYMES
EVEN-ODD NUCLEI
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
NITROGEN TRANSFERASES
NUCLEI
RADIOISOTOPES
REACTION KINETICS
SEPARATION PROCESSES
SULFUR ISOTOPES
TRANSFERASES
550201* - Biochemistry- Tracer Techniques