Proteomic study of caveolae and rafts isolated from human endothelial cells

Richard R Sprenger, Anton J G Horrevoets

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Caveolae and rafts are specialized microdomains of the endothelial cell plasma membrane, which play an important role in signal transduction, transcellular transport, and cholesterol homeostasis. The dynamic protein composition of these subcellular lipid domains has been implicated in a variety of patho-physiological states of the vasculature, and is receiving increased attention. As a result of the membranous composition and abundance of insoluble intrinsic and membrane-associated proteins, determination of the raft/caveolae subproteome composition requires specially adapted methods. In this chapter, we present a straightforward protocol to obtain comprehensive and reliable peptide mixtures from this subproteome by subcellular fractionation and both one-dimensional and two-dimensional gel electrophoresis. These mixtures allow dynamic monitoring of composition and posttranslational modification of the raft/caveola subproteome using peptide mass fingerprinting and direct peptide sequencing tandem mass spectrometry.
Original languageEnglish
Book seriesMethods in Molecular Biology
Volume357
Pages (from-to)199-213
Number of pages14
ISSN1064-3745
DOIs
Publication statusPublished - 1. Jan 2007
Externally publishedYes

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