Comparative proteomic analysis of a cytosolic fraction from β3 integrin-deficient cells

Jason A. Bush, Hideki Kitaura, Yuliang Ma, Steven L. Teitelbaum, F. Patrick Ross, Jeffrey W. Smith

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Integrins are heterodimeric transmembrane receptors involved in sensing and transmitting informational cues from the extracellular environment to the cell. This study explored sub-proteome changes in response to elimination of the β3 integrin using a knockout murine model. Cleavable isotope-coded affinity tagging (cICAT) in combination with sub-cellular fractionation, multiple dimensions of separation and tandem mass spectrometry (MS/MS) were used to characterize differentially expressed proteins among β3 integrin -/- (β3 -/- ) mouse embryonic fibroblasts and isogenic wild-type (WT) controls. From a cytosolic protein fraction, 48 proteins were identified, in which expression differed by >1.5-fold. Predominant ontological groups included actin-binding/cytoskeletal proteins and protease/protease inhibitors. Interestingly, β3 integrin expression was inversely correlated with expression of cathepsin B, a lysosomal cysteine protease, as its expression was greater by over 3.5-fold in the β3 -/- cells. This inverse correlation was also observed in stable heterologous cells transfected with β3 integrin, where the intracellular expression and activity of cathepsin B was lower compared to control cells. Our data suggests that the composition of the cellular proteome is influenced by integrin expression patterns and reveals a strong functional relationship between β3 integrin and cathepsin B.

Original languageEnglish
Pages (from-to)1-13
Number of pages13
JournalCancer Genomics and Proteomics
Volume9
Issue number1
Publication statusPublished - 2012 Jan 1
Externally publishedYes

Keywords

  • Cancer-proteomics
  • Cathepsin B
  • Fractionation
  • GeLC-MS
  • HEK293 cells
  • Integrin
  • Isotope-coded affinity tags
  • Protease

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cancer Research

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