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    Enzymatic Glyco-Modification of Synthetic Membrane Systems

    Year: 2023

    Authors: Dylan Jabeguero, Lina Siukstaite, Chunyue Wang, Anna Mitrovic,Serge Pérez,Olga Makshakova,Ralf P. Richter,Winfried Römer and Christelle Breton

    Keywords: giant unilamellar vesicles; globotriaosylceramide; LgtC; molecular dynamics; supported lipid bilayer

    The present report assesses the capability of a soluble glycosyltransferase to modify glycolipids organized in two synthetic membrane systems that are attractive models to mimic cell membranes: giant unilamellar vesicles (GUVs) and supported lipid bilayers (SLBs). The objective was to synthesize the Gb3 antigen (Galα1,4Galβ1,4Glcβ-Cer), a cancer biomarker, at the surface of these membrane models. A soluble form of LgtC that adds a galactose residue from UDP-Gal to lactose-containing acceptors was selected. Although less efficient than with lactose, the ability of LgtC to utilize lactosyl–ceramide as an acceptor was demonstrated on GUVs and SLBs. The reaction was monitored using the B-subunit of Shiga toxin as Gb3-binding lectin. Quartz crystal microbalance with dissipation analysis showed that transient binding of LgtC at the membrane surface was sufficient for a productive conversion of LacCer to Gb3. Molecular dynamics simulations provided structural elements to help rationalize experimental data.
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