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Radioisotopic assay of picomolar amounts of coenzyme A

✍ Scribed by Daniel Rabier; Pascale Briand; Françoise Petit; Pierre Kamoun; Liliane Cathelineau

Publisher
Elsevier Science
Year
1983
Tongue
English
Weight
420 KB
Volume
134
Category
Article
ISSN
0003-2697

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✦ Synopsis

A two-step method of determining reduced coenzyme A (CoASH) concentrations in tissue or cell extracts is described. In the first step, CoASH is reacted with acetylphosphate in a reaction catalyzed by phosphotransacetylase to yield acetyl-CoA. Acetyl-CoA is then condensed with [?]oxaloacetate by citrate synthase to give ["'Clcitrate. This method allows the measurement of lo-200 pmol of CoASH. By omitting the phospbotransacetylase step, measurement of the same amount of acetyl-CoA is possible. ' Abbreviations used: CoASH, reduced Coenzyme A; GOT, glutamate-oxaloacetate transaminase; NEM, Nethylmaleimide. MATERIALS AND METHODS Enzymes. Phosphotransacetylase (Clostridium kluyveri, lyophilized powder, 220 U/mg) 325 0003~2697/83 $3.00 C opvnght 'C' 19X3 hy Acadcm~c I'm,. fnr ,I, nghrs d rcprduclm I" 311,' ii,,", rc5cnc<,

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