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Membrane remodeling capacity of a vesicle-inducing glycosyltransferase

par Ge, Changrong ;Gõmez-Llobregat, Jordi;Skwark, Marcin M.J.;Wieslander, Åke A.;Lindén, Martin;Ruysschaert, Jean Marie
Référence The FEBS journal, 281, 16, page (3667-3684)
Publication Publié, 2014

Article révisé par les pairs

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Résumé :

Intracellular vesicles are abundant in eukaryotic cells but absent in the Gram-negative bacterium Escherichia coli. However, strong overexpression of a monotopic glycolipid-synthesizing enzyme, monoglucosyldiacylglycerol synthase from Acholeplasma laidlawii (alMGS), leads to massive formation of vesicles in the cytoplasm of E. coli. More importantly, alMGS provides a model system for the regulation of membrane properties by membrane-bound enzymes, which is critical for maintaining cellular integrity. Both phenomena depend on how alMGS binds to cell membranes, which is not well understood. Here, we carry out a comprehensive investigation of the membrane binding of alMGS by combining bioinformatics methods with extensive biochemical studies, structural modeling and molecular dynamics simulations. We find that alMGS binds to the membrane in a fairly upright manner, mainly by residues in the N-terminal domain, and in a way that induces local enrichment of anionic lipids and a local curvature deformation. Furthermore, several alMGS variants resulting from substitution of residues in the membrane anchoring segment are still able to generate vesicles, regardless of enzymatic activity. These results clarify earlier theories about the driving forces for vesicle formation, and shed new light on the membrane binding properties and enzymatic mechanism of alMGS and related monotopic GT-B fold glycosyltransferases. We characterize the membrane binding properties of the monotopic glycosyltransferase alMGS, and find that it binds in an upright manner and induces local enrichment of anionic lipids and local curvature deformations. These results shed new light on the ability of alMGS to induce intracellular vesicles upon overexpression, and on the enzymatic mechanism of alMGS and related GT-B fold glycosyltransferases. © 2014 FEBS.