A novel strategy for the comprehensive analysis of the biomolecular composition of isolated plasma membranes.
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.
Department of Molecular and Developmental Genetics (VIB11), Laboratory for Membrane Trafficking and Center for Human Genetics (KULeuven), Gasthuisberg O&N4, Leuven, Belgium.
Deepak B
DB
Thimiri Govinda Raj
Bart
B
Ghesquière
Arun Kumar
AK
Tharkeshwar
Katrijn
K
Coen
Rita
R
Derua
Dieter
D
Vanderschaeghe
Evelien
E
Rysman
Murali
M
Bagadi
Pieter
P
Baatsen
Bart
B
De Strooper
Etienne
E
Waelkens
Gustaaf
G
Borghs
Nico
N
Callewaert
Johan
J
Swinnen
Kris
K
Gevaert
Wim
W
Annaert
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000032:analysis / Q000378:metabolism; D011992:Endosomes / Q000235:genetics / Q000473:pathology; D005347:Fibroblasts; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D058185:Magnetite Nanoparticles; D008563:Membrane Lipids / Q000032:analysis / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D051379:Mice; D053763:Presenilins / Q000235:genetics*; D040901:Proteomics / Q000379:methods*; D013109:Sphingomyelins / Q000032:analysis / Q000378:metabolism
D058185:Magnetite Nanoparticles; D008563:Membrane Lipids; D008565:Membrane Proteins; D053763:Presenilins; D013109:Sphingomyelins; D002784:Cholesterol