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Plasma membrane‐focused proteomics: Dramatic changes in surface expression during the maturation of human dendritic cells
Author(s) -
Watarai Hiroshi,
Hinohara Atsushi,
Nagafune Jun,
Nakayama Toshinori,
Taniguchi Masaru,
Yamaguchi Yasunori
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401258
Subject(s) - microbiology and biotechnology , proteomics , transmembrane protein , membrane protein , chemistry , cell , biology , membrane , biochemistry , receptor , gene
The differential expression of surface molecules on dendritic cells (DC) reflects their functional differences as immature and mature subsets. It is difficult, however, to characterize differences in surface expression by standard proteomic approaches, due mainly to the hydrophobic nature and low abundance of the individual proteins in question. We have established a method for obtaining high‐yield plasmalemma preparations which contain surface molecules enriched more than 200‐fold by coating cells with beads conjugated with antibody against a cell type‐specific cell‐surface molecule, followed by nitrogen cavitated disruption, magnetic separation, and density gradient ultracentrifugation. We identified and quantified 339 human monocyte‐derived DC transmembrane proteins, including 33 previously uncharacterized molecules. Whereas 106 proteins were selectively expressed in immature cells or down‐regulated after maturation, 191 proteins were selectively expressed in mature cells or up‐regulated after maturation.