Dynamic modulation of the glycosphingolipid content in supported lipid bilayers by glycolipid transfer protein.
Supported lipid bilayers (SLBs) are popular models of cell membranes. Owing to the importance of glycosphingolipids (GSLs) in modulating structure and function of membranes and membrane proteins, methods to tune the GSL content in SLBs would be desirable. Glycolipid transfer protein (GLTP) can selectively transfer GSLs between membrane compartments. Using the ganglioside GM1 as a model GSL, and two mass-sensitive and label-free characterization techniques-quartz crystal microbalance with dissipation monitoring and ellipsometry-we demonstrate that GLTP is an efficient and robust biochemical tool to dynamically modulate the GSL content of SLBs up to 10 mol % GM1, and to quantitatively control the GSL content in the bulk-facing SLB leaflet. By exploiting what we believe to be a novel tool, we provide evidence that GM1 distributes highly asymmetrically in silica-supported lipid bilayers, with ∼85% of the ganglioside being present in the bulk-facing membrane leaflet. We report also that the pentameric B-subunit of cholera toxin binds with close-to-maximal stoichiometry to GM1 in SLBs over a large range of GM1 concentrations. Furthermore, we quantify the liganding affinity of GLTP for GM1 in an SLB context to be 1.5 μM.
Biosurfaces Unit, Cooperative Research Center in Biomaterials (CIC biomaGUNE), San Sebastian, Spain.
Ixaskun
I
Carton
Lucy
L
Malinina
Ralf P
RP
Richter
D055592:Biophysical Phenomena; D002352:Carrier Proteins / Q000737:chemistry*; D002772:Cholera Toxin / Q000737:chemistry; D005677:G(M1) Ganglioside / Q000737:chemistry; D006028:Glycosphingolipids / Q000737:chemistry*; D066298:In Vitro Techniques; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry*; D008954:Models, Biological; D058525:Quartz Crystal Microbalance Techniques; D011994:Recombinant Proteins / Q000737:chemistry
D002352:Carrier Proteins; D006028:Glycosphingolipids; D008024:Ligands; D008051:Lipid Bilayers; D011994:Recombinant Proteins; C030363:lipid transfer protein; D005677:G(M1) Ganglioside; D002772:Cholera Toxin