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

Molecular Systems Biology

Deepak B Thimiri Govinda Raj⁷  Bart Ghesquière⁵  Arun Kumar Tharkeshwar⁷  Katrijn Coen⁷  Rita Derua³  Dieter Vanderschaeghe⁶  Evelien Rysman¹  Murali Bagadi¹  Pieter Baatsen⁴  Bart De Strooper⁸  Etienne Waelkens³  Gustaaf Borghs²  Nico Callewaert⁶  Johan Swinnen¹  Kris Gevaert⁵ 

[1]Department of Experimental Medicine and Endocrinology (KULeuven), Leuven, Belgium

[2]Department of Bio-Nano Electronics, Functional Nanosystems group, Interuniversity Microelectronics Center (IMEC), Leuven, Belgium

[3]Department of Molecular Cell Biology (KULeuven), Leuven, Belgium

[4]Department of Molecular and Developmental Genetics (VIB11 and KULeuven), Electron Microscopy Network (EMoNe), Leuven, Belgium

[5]Department of Medical Protein Research, VIB and Department of Biochemistry, UGent, Ghent, Belgium

[6]Department of Molecular Biomedical Research, Unit for Medical Biotechnology, VIB and UGent, Ghent, Belgium

[7]Department of Molecular and Developmental Genetics (VIB11), Laboratory for Membrane Trafficking and Center for Human Genetics (KULeuven), Leuven, Belgium

[8]Department of Molecular and Developmental Genetics (VIB11), Laboratory for Neurodegenerative Diseases and Center for Human Genetics (KULeuven), Leuven, Belgium

关键词: eukaryotic cell systems;    glycomics;    lipidomics;    presenilin;    proteomics;   

DOI  :  10.1038/msb.2011.74

来源: Wiley

【 摘 要 】

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 P

【 授权许可】

CC BY-NC-SA   
Copyright © 2011 EMBO and Macmillan Publishers Limited

Creative Commons Attribution License, which permits distribution, and reproduction in any medium, provided the original author and source are credited. This license does not permit commercial exploitation without specific permission.