title: An improved method for the assay of platelet pyruvate dehydrogenase.
abstract: An improved method for the assay of human platelet pyruvate dehydrogenase is described. By generating the substrate [1-14C]pyruvate in situ from [1-14C]lactate plus L-lactate dehydrogenase, the rate of spontaneous decarboxylation is dramatically reduced, allowing far greater sensitivity in the assay of low activities of pyruvate dehydrogenase. In addition, no special precautions are required for the storage and use of [1-14C]lactate, in contrast to those for [1-14C]pyruvate. These factors allow a 5-10-fold increase in sensitivity compared with current methods. The pyruvate dehydrogenase activity of normal subjects as determined by the [1-14C]lactate system was 215 +/- 55 pmol . min-1 . mg-1 protein (n = 18). The advantages of this assay system are discussed.
mesh_terms: D000328:Adult; D001774:Blood Chemical Analysis / Q000379:methods; D001792:Blood Platelets / Q000201:enzymology*; D001793:Blood Preservation; D003653:Decarboxylation; D006801:Humans; D011768:Pyruvate Dehydrogenase Complex / Q000097:blood*
chemical_list: D011768:Pyruvate Dehydrogenase Complex