Global profiling of the cell surface proteome of cancer cells uncovers an abundance of proteins with chaperone function

J Biol Chem. 2003 Feb 28;278(9):7607-16. doi: 10.1074/jbc.M210455200. Epub 2002 Dec 18.

Abstract

There is currently limited data available pertaining to the global characterization of the cell surface proteome. We have implemented a strategy for the comprehensive profiling and identification of surface membrane proteins. This strategy has been applied to cancer cells, including the SH-SY5Y neuroblastoma, the A549 lung adenocarcinoma, the LoVo colon adenocarcinoma, and the Sup-B15 acute lymphoblastic leukemia (B cell) cell lines and ovarian tumor cells. Surface membrane proteins of viable, intact cells were subjected to biotinylation then affinity-captured and purified on monomeric avidin columns. The biotinylated proteins were eluted from the monomeric avidin columns as intact proteins and were subsequently separated by two-dimensional PAGE, transferred to polyvinylidene difluoride membranes, and visualized by hybridization with streptavidin-horseradish peroxidase. Highly reproducible, but distinct, two-dimensional patterns consisting of several hundred biotinylated proteins were obtained for the different cell populations analyzed. Identification of a subset of biotinylated proteins among the different cell populations analyzed using matrix-assisted laser desorption ionization and tandem mass spectrometry uncovered proteins with a restricted expression pattern in some cell line(s), such as CD87 and the activin receptor type IIB. We also identified more widely expressed proteins, such as CD98, and a sushi repeat-containing protein, a member of the selectin family. Remarkably, a set of proteins identified as chaperone proteins were found to be highly abundant on the cell surface, including GRP78, GRP75, HSP70, HSP60, HSP54, HSP27, and protein disulfide isomerase. Comprehensive profiling of the cell surface proteome provides an effective approach for the identification of commonly occurring proteins as well as proteins with restricted expression patterns in this compartment.

MeSH terms

  • Activin Receptors, Type II / metabolism
  • Amino Acid Sequence
  • Biotinylation
  • Blotting, Western
  • Carrier Proteins / biosynthesis
  • Electrophoresis, Gel, Two-Dimensional
  • Endoplasmic Reticulum Chaperone BiP
  • Fusion Regulatory Protein-1 / biosynthesis
  • HSP70 Heat-Shock Proteins / biosynthesis
  • Heat-Shock Proteins*
  • Humans
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Molecular Chaperones / biosynthesis
  • Molecular Chaperones / metabolism*
  • Molecular Sequence Data
  • Neoplasms / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Protein Array Analysis
  • Protein Binding
  • Protein Structure, Tertiary
  • Proteome
  • Receptors, Cell Surface / biosynthesis
  • Receptors, Urokinase Plasminogen Activator
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tumor Cells, Cultured

Substances

  • Carrier Proteins
  • Endoplasmic Reticulum Chaperone BiP
  • Fusion Regulatory Protein-1
  • HSP70 Heat-Shock Proteins
  • HSPA5 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • PLAUR protein, human
  • Proteome
  • Receptors, Cell Surface
  • Receptors, Urokinase Plasminogen Activator
  • Activin Receptors, Type II
  • activin receptor type II-B