A new phosphoglycerolipid, 'phosphatidylglucose', found in human cord red cells by multi-reactive monoclonal anti-i cold agglutinin, mAb GL-1/GL-2

Cord red cell membranes express many differentiation-related molecules. To study such molecules, we have established human cell lines, termed CL-1 and GL-2, by the Epstein-Barr virus transformation method, both of which produ ce monoclonal anti-i cold agglutinin [Y. Nagatsuka et al,, Immunol. Lett, 4 6 (1995) 93-100], Thin layer chromatography immunoblotting analysis reveale d that these antibodies had broad specificities reacting with a variety of glycolipid antigens. Of the immunoreactive lipid antigens, a new phosphogly cerolipid containing glucose from human cord red cells was found. The isola ted lipid was unstable to alkaline hydrolysis and contained glucose as a so le sugar, Secondary ion mass spectrum-collision-induced dissociation mass s pectrometric analysis of this lipid gave the main molecular ion peak at m/z 885 corresponding to phosphatidylhexose. This antigen was susceptible to p hospholipases A2, C and D but resistant to phosphatidylinositol-specific ph ospholipase C. Two-dimensional nuclear magnetic resonance spectroscopy conf irmed that glucose is linked to the sn-glycerol 3-phosphate residue with a p-anomeric configuration, Based upon these combined results, we identified this lipid as phosphatidyl-beta -D-glucose. This is the first report showin g the presence of the glucosylated glycerophospholipid in mammalian sources . (C) 2001 Federation of European Biochemical Societies. Published by Elsev ier Science B.V. All rights reserved.