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Phospholipase D hydrolyzes short-chain analogs of phosphatidylcholine in the absence of detergent
Journal article   Open access   Peer reviewed

Phospholipase D hydrolyzes short-chain analogs of phosphatidylcholine in the absence of detergent

Leslee Davis, Jeffrey Maglio and Joel Horwitz
Lipids, v 33(2)
Feb 1998
PMID: 9507245
url
https://doi.org/10.1007/s11745-998-0199-5View
Published, Version of Record (VoR)Maybe Open Access (Publisher Bronze) Open

Abstract

Life Sciences Medicinal Chemistry Biochemistry, general Nutrition Microbial Genetics and Genomics Medical Biochemistry Bioorganic Chemistry
Phospholipase D is an important enzyme in signal transduction in neuronal tissue. A variety of assays have been used to measure phospholipase D activity in vitro. The most typical measure of phospholipase D activity is the production of phosphatidylethanol in the presence of ethanol. Phosphatidylethanol is a product of transphosphatidylation activity that is considered a unique property of phospholipase D. To support transphosphatidylation activity, high concentrations of ethanol may be required. Furthermore, most assays in the literature utilize a detergent. These extreme conditions, detergent and ethanol, may alter phospholipase D and hinder the study of its regulation. In this manuscript we describe an assay that eliminates these potentially confounding conditions. It utilizes high specific activity [3H]butanol as a nucleophilic receptor. This eliminates the need for high concentrations of alcohol. The substrate is an analog of phosphatidylcholine that contains short-chain fatty acids, 1,2-dioctanoyl-sn-glycero-3-phosphocholine. Phospholipase D readily hydrolyzes this substrate in the absence of detergent. This novel assay should be useful in the further characterization of phospholipase D.

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Web of Science research areas
Biochemistry & Molecular Biology
Nutrition & Dietetics
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