Analysis of cell surface proteome changes via label-free, quantitative mass spectrometry.
We present a mass spectrometry-based strategy for the specific detection and quantification of cell surface proteome changes. The method is based on the label-free quantification of peptide patterns acquired by high mass accuracy mass spectrometry using new software tools and the cell surface capturing technology that selectively enriches glycopeptides exposed to the cell exterior. The method was applied to monitor dynamic protein changes in the cell surface glycoproteome of Drosophila melanogaster cells. The results led to the construction of a cell surface glycoprotein atlas consisting of 202 cell surface glycoproteins of D. melanogaster Kc167 cells and indicated relative quantitative changes of cell surface glycoproteins in four different cellular states. Furthermore we specifically investigated cell surface proteome changes upon prolonged insulin stimulation. The data revealed insulin-dependent cell surface glycoprotein dynamics, including insulin receptor internalization, and linked these changes to intracellular signaling networks.
Schiess
Ralph
R
Institute of Molecular Systems Biology, Eidgenössische Technische Hochschule (ETH) Zurich, 8093 Zurich, Switzerland.
Mueller
Lukas N
LN
Schmidt
Alexander
A
Mueller
Markus
M
Wollscheid
Bernd
B
Aebersold
Ruedi
R
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002853:Chromatography, Liquid; D029721:Drosophila Proteins / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology* / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects; D006023:Glycoproteins / Q000032:analysis; D007328:Insulin / Q000494:pharmacology; D007553:Isotope Labeling; D013058:Mass Spectrometry / Q000379:methods*; D020543:Proteome / Q000032:analysis*; D011972:Receptor, Insulin / Q000378:metabolism; D015203:Reproducibility of Results
D029721:Drosophila Proteins; D006023:Glycoproteins; D007328:Insulin; D020543:Proteome; D011972:Receptor, Insulin