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A novel strategy for the comprehensive analysis of the biomolecular composition of isolated plasma membranes.

Molecular systems biology (2011-10-27)
Deepak B Thimiri Govinda Raj, Bart Ghesquière, Arun Kumar Tharkeshwar, Katrijn Coen, Rita Derua, Dieter Vanderschaeghe, Evelien Rysman, Murali Bagadi, Pieter Baatsen, Bart De Strooper, Etienne Waelkens, Gustaaf Borghs, Nico Callewaert, Johan Swinnen, Kris Gevaert, Wim Annaert
ABSTRACT

We manufactured a novel type of lipid-coated superparamagnetic nanoparticles that allow for a rapid isolation of plasma membranes (PMs), enabling high-resolution proteomic, glycomic and lipidomic analyses of the cell surface. We used this technology to characterize the effects of presenilin knockout on the PM composition of mouse embryonic fibroblasts. We found that many proteins are selectively downregulated at the cell surface of presenilin knockout cells concomitant with lowered surface levels of cholesterol and certain sphingomyelin species, indicating defects in specific endosomal transport routes to and/or from the cell surface. Snapshots of N-glycoproteomics and cell surface glycan profiling further underscored the power and versatility of this novel methodology. Since PM proteins provide many pathologically relevant biomarkers representing two-thirds of the currently used drug targets, this novel technology has great potential for biomedical and pharmaceutical applications.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
1,2-Hexadecanediol, technical grade, 90%