title: Detection of in situ labeled cell surface proteins by mass spectrometry: application to the membrane subproteome of human mammary epithelial cells.
abstract: Characterization of the surface exposed membrane subproteome of human mammary epithelial cells (strain 184 A1L5) implemented lysine specific in situ labeling of the proteins using sulfosuccinimidyl-6-(biotinamido)hexanoate, followed by enrichment of the biotinylated, tryptically digested peptides, and then liquid chromatography-tandem mass spectrometry analysis of the labeled peptides. Probing the membrane subproteome in this manner yielded unambiguous identification of proteins situated on the cell surface. The method reported can be adapted to include stable isotope labeling of proteins for quantitation of changes occurring on the cell surface in response to specific perturbations.
authors.affiliation: Cell Biology, Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors.forename: Wan-Nan U
authors.initials: WN
authors.lastname: Chen
authors.forename: Li-Rong
authors.initials: LR
authors.lastname: Yu
authors.forename: Eric F
authors.initials: EF
authors.lastname: Strittmatter
authors.forename: Brian D
authors.initials: BD
authors.lastname: Thrall
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D004847:Epithelial Cells / Q000737:chemistry; D008321:Mammary Glands, Animal / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D040901:Proteomics
chemical_list: D008565:Membrane Proteins