出版社：American Society for Biochemistry and Molecular Biology
摘要：A procedure for stereoanalysis of radiochemically labeled glycerophospholipids is described. It is based on the study of the labeled alpha-glycerophosphate which retains its original configuration when liberated upon alkaline hydrolysis of the lipids. The labeled alpha-glycerophosphate is oxidized enzymatically with sn-3-glycerophosphate dehydrogenase and the product, dihydroxyacetone phosphate, is degraded with alkali to inorganic phosphate. The nonoxidizable alpha-glycerophophate (sn-1-glycerophosphate), the beta-glycerophosphate, and the inorganic phosphate derived from sn-3-glycerophosphate are quantitated after separation by thin-layer chromatography. The procedure gave the expected results when applied to [3H]glycerol-and 32P-labeled phosphatidylcholine, bis( monoacylglycero)phosphate, and phosphatidylglycerol from natural resources. Bis(monoacylglycero)phosphate, known also as lysobisphosphatidic acid, was synthesized from ]32P]diphosphatidylglycerol and from phosphatidyl[1',3'-3H]glycerol in lysosomal preparations of rat liver according to Poorthuis and Hostetler (1978. J. Lipid Res. 19: 309-315). Stereoanalysis proved that the product was in both cases a derivate of sn-1-glycerophospho-sn-1'-glycerol.
