Correlation between bilayer lipid dynamics and activity of the diglucosyldiacylglycerol synthase from Acholeplasma laidlawii membranes.

In the single membrane of Acholeplasma laidlawii a specific glucosyltransferase synthesize the major, lamellar-forming lipid diglucosyldiacylglycerol (DGlcDAG) from the major, nonlamellar-prone monoglucosyldiacylglycerol (MGlcDAG). This is crucial for the maintenance of phase equilibria close to a bilayer-nonbilayer transition and a nearly constant spontaneous curvature in the membrane lipid bilayer. Acyl chain order is also affected, but not kept constant. Phosphatidylglycerol (PG) is an essential activator, needed in substantial amounts by the DGlcDAG synthase, and likely to affect bilayer properties. A potential connection was investigated between the (i) lateral diffusion, (ii) domain formation of the PG activator and (iii) bilayer chain ordering (i.e., the hydrocarbon free volume), revealed in unilamellar liposomes by lipid probes containing one or two (fluorescent) pyrene acyl chains, and (iv) activity of the DGlcDAG synthase. Different activator, nonbilayer perturbant, and bilayer matrix conditions were employed. Diffusion of PG was substantially slower in a DGlcDAG compared to a phosphatidylcholine (PC) matrix with 18:1c chains but increased with the PG content in both. No obvious correlation between diffusion and enzyme activity, and no local concentration of PG as a function of chain ordering or curvature, was detected. However, an enrichment of PG activator into domains could be induced by a chain length mismatch between 18:1c-PG and 14:1c-PC (but not 22:1c-PC), even at small PG fractions. Patching was sufficient to stimulate enzyme activity 4-fold in relation to the activities normally valid at low PG concentrations. Chain order was substantially lower (i.e., free volumes larger) in bilayers of DGlcDAG than in bilayers of PC and increased in an additive fashion in both by the content of especially the nonbilayer-prone 1,3-18:1c-DAG but also by PG. At physiological concentrations of PG in DGlcDAG bilayers (approximately 20%) a good correlation was evident between increased DAG content and chain ordering and strongly enhanced enzyme activities, with maxima close to a bilayer-nonbilayer transition. It is concluded that regulation of packing conditions in A. laidlawii membranes by the DGlcDAG synthase seems to be governed not by the absolute extent of chain order but more by the spontaneous curvature within a certain range of conditions. Domain formation of the essential PG activator due to bilayer conditions is a second mechanism, potentially overriding the curvature effects.