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Metabolism and Enzymology

Separation and Characterization of Inositol Phospholipids from the Pulvini of Samanea saman¹

Department of Molecular and Cell Biology U-125, University of Connecticut, Storrs, Connecticut 06269

To supplement current thin-layer chromatographic methods for separation and quantitation of plant phospholipids, an alternative method, high-performance liquid chromatography was developed. The major inositol-containing lipids from the pulvini of Samanea saman Merr. were identified as phosphatidylinositol, phosphatidylinositol phosphate, and phosphatidylinositol bisphosphate based on comigration with authentic standards on high-performance liquid chromatography and on thin-layer chromatography. The patterns of incorporation of radioactivity into the putative phosphatidylinositol and phosphatidylinositol phosphate were consistent with these identifications when pulvini were labeled with [³H]glycerol, [³H]inositol, or [³²P]orthophosphate. Analysis of the products of enzymic hydrolysis, of chemical deacylation, and of `fingerprint' methanolysis of these phospholipids confirmed the identifications.

³ Present address: Department of Botany, Connecticut College, New London, CT 06320.

¹ This work was supported by National Science Foundation (NSF) grant DMB 86-07857 to R. C. C. and R. L. S., and by NSF grants DMB 83-04613 and DCB 88-04810 to R. L. S.

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