A spectrophotometric method for the assay of phospholipase D activity

George M. Carman, Anthony S. Fischl, Maureen Dougherty, Gerhard Maerker

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Phosphatidylcholine phosphatidohydrolase (EC 3.1.4.4, phospholipase D) catalyzes the hydrolysis of phosphatidylcholine to phosphatidic acid and choline. We have developed a spectrophotometric assay for phospholipase D using choline kinase, pyruvate kinase, and lactate dehydrogenase to couple the release of choline with the oxidation of NADH. The assay was linear both with time and with enzyme concentration. The assay should prove useful for continuous monitoring of enzyme activity, determination of initial rates of reaction, and detailed kinetic studies of phospholipase D. The method is limited to analysis of purified preparations of phospholipase D lacking competing activities to the coupled system.

Original languageEnglish (US)
Pages (from-to)73-76
Number of pages4
JournalAnalytical Biochemistry
Volume110
Issue number1
DOIs
StatePublished - Jan 1 1981

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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