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PubMed1983

A method for the synthesis of D-3-phospho[U-14C]glycerate.

An enzymatic method for the synthesis of radioactive D-3-phosphoglycerate from commercially available D-[U-14C]fructose 1,6-diphosphate is described. The unique aspect of this procedure is the substitution of arsenate for phosphate in the glyceraldehyde-3-phosphate dehydrogenase reaction. The 1-arseno-3-phosphoglycerate formed spontaneously hydrolyzes to form the D-3-phosphoglycerate product. The methods detailed below for the synthesis, isolation, and analysis of the 3-phospho[U-14C]glycerate product are relatively easy.

Carbon Radioisotopes↗
PubMed1985

Determination of picomole amounts of glycerate 3-phosphate, glycerate 2-phosphate, and phosphoenol pyruvate by an enzymatic assay coupled to firefly luciferase/luciferin luminescence.

A procedure for the determination of picomole amounts of glycerate 3-phosphate, glycerate 2-phosphate, and phosphoenol pyruvate is described. These metabolites were utilized by the glycolytic enzymes phosphoglycerate mutase, enolase, and pyruvate kinase to generate ATP which was determined by firefly luciferase/luciferin luminescence. The phosphoglycerate mutase used was of the glycerate 2,3-bisphosphate-independent type and was prepared from wheat germ. Stoichiometric conversion of glycerate 3-P, ranging in amount from 9 to 275 pmol, occurred after 25 min preincubation and required a narrow range of added mutase. The application of the procedure for determining these metabolites in suspensions of plant protoplasts is described.

Adenosine Diphosphate↗