The structures of two phosphoglucolipids from the membrane of Acholeplasma laidlawii, strain A-EF22 were determined by high resolution 13C-, 31P- and 1H-NMR. The lipids in question are 1,2-diacyl-3-O-[glycerophosphoryl-6-O-(alpha-D-glucopyranosyl- (1-->2)-O-alpha-D-glucopyranosyl)]-sn-glycerol (1) and 1,2-diacyl-3-O-[glycerophosphoryl-6-O-(alpha-D-glucopyranosyl-(1-- >2)- monoacyl-glycerophosphoryl-6-O-alpha-D-glucopyranosyl)]-sn-glycero l (2). Both lipids are thus derivatives of diglucosyldiacylglycerol. Previous reports on these lipids, based on insufficient chemical analyses, showed contradictory structures. A phosphoglycolipid having the structure of 2 has not been described previously.