New EPR method for cellular surface characterization.
An electron paramagnetic resonance (EPR)-based membrane surface characterization method is presented to detect the properties of the carbohydrate-rich part of membrane surfaces as well as carbohydrate interaction with other membrane constituents and water-soluble molecules. The proposed method relies on the spin-labeling and spectral decomposition based on spectral simulation and optimization with EPRSIM software. In order to increase the sensitivity of characterization to the carbohydrate-rich part of the membrane surface, the sucrose-contrasting approach is introduced. With this method, which was established on model membranes with glycolipids and tested on erythrocyte membrane, we were able to characterize the surface and lipid bilayer lateral heterogeneity. Additionally, some properties of the interaction between glycocalyx and lipid bilayer as well as between glycocalyx and sucrose molecules were determined. The experiments also provided some information about the anchoring and aggregation of the glycosylated molecules. According to the results, some functions of the glycosylated surface are discussed.
Strancar
J
J
Jozef Stefan Institute, Jamova 39, SI-1000 Ljubljana, Slovenia. janez.strancar@ijs.si
Schara
M
M
Pecar
S
S
D000465:Algorithms; D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D004910:Erythrocyte Membrane / Q000737:chemistry*; D005732:Gangliosides / Q000737:chemistry*; D019276:Glycocalyx / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances; D008560:Membrane Fluidity; D021962:Membrane Microdomains / Q000737:chemistry; D013113:Spin Labels; D013395:Sucrose / Q000737:chemistry*; D013499:Surface Properties
D002241:Carbohydrates; D005732:Gangliosides; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D013113:Spin Labels; D013395:Sucrose