Hydrolysis of N-acylated glycerophospholipids by phospholipases A2 and D: A method of identification and analysis

✍ Scribed by P.C. Schmid; V. Natarajan; B.K. Weis; H.H.O. Schmid

• Book ID: 103038810
• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 628 KB
• Volume: 41
• Category: Article
• ISSN: 0009-3084
• DOI: 10.1016/0009-3084(86)90022-8

✦ Synopsis

We have previously identified N-acylethanolamine phospholipids in infarcted dog heart and in normal fish brain by chemical and enzymatic degradation. We now report that hydrolysis with phospholipase D from Streptomyces chromofuscus removes N-acylethanolamine from N-acylethanolamine phospholipids and lyso N-acylethanolamine phospholipids, or N-acylserine from lyso N-acylserine phospholipids. At acidic pH, a phosphatase present in the phospholipase D preparation further hydrolyzes the resulting phosphatidic acid (PA) or lyso-PA to diacyl-or monoacylglycerol. Because N-acylserine phospholipids are a poor substrate for the phospholipase D, pretreatment with phospholipase A 2 (Trimeresurus flavoviridis venom) is used to remove the 2-O-acyl group. Thus, both types of N-acylated phospholipids can be analyzed by consecutive phospholipase A 2 and phospholipase D treatment. Reaction products, i.e., free fatty acids, monoacylglycerols and N-acylethanolamine or N-acylserine, are separable by thinlayer chromatography. Both N-acyl components can be further characterized by conversion to the t-butyldimethylsilyl derivatives. The method was used to identify and analyze the Nacylserine phospholipids of bovine brain.