Plasma membrane-focused proteomics: dramatic changes in surface expression during the maturation of human dendritic cells.
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.
Watarai
Hiroshi
H
Laboratory for Immune Regulation, RIKEN Research Center for Allergy and Immunology, Yokohama, Japan. hwatarai@rcai.riken.jp
Hinohara
Atsushi
A
Nagafune
Jun
J
Nakayama
Toshinori
T
Taniguchi
Masaru
M
Yamaguchi
Yasunori
Y
D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology; D040901:Proteomics
D008565:Membrane Proteins