Asymmetrical topology of diether- and tetraether-type polar lipids in membranes of Methanobacterium thermoautotrophicum cells.

We investigated the distribution of diether polar lipids between the inner and outer leaflets of the membrane of Methanobacterium thermoautotrophicum comparing the orientation of tetraether polar lipids, which constitute a monolayer in the same membrane. Three kinds of reactions were employed for intact cells or protoplasts and unsealed membrane fragments prepared from the organism: glycosidase digestion for glycolipids, NaIO4 oxidation for glycolipids and inositol lipids, and trinitrophenylation for aminophospholipids. The results indicated that (a) most gentiobiose residues of both diether and tetraether polar lipids were exposed on the outside of the cells; (b) serine and inositol residues of both diether and tetraether polar lipids were mainly oriented to the cytoplasmic surface of the membrane; and (c) approximately 80% of archaetidylethanolamine (diether type) was distributed in the outer leaflet of the membrane bilayer, while only 25% of the ethanolamine residue of gentiobiosyl caldarchaetidylethanolamine (tetraether type) was oriented to the outer surface of the membrane. These results, except for ethanolamine lipids, are consistent with the hypothesis that the tetraether polar lipids are synthesized from the corresponding diether polar lipid precursors that have been already substituted by polar groups in the membrane by head-to-head condensation without rearrangement of lipids.