Proteomic profiling identifies breast tumor metastasis-associated factors in an isogenic model

Paweena Kreunin, Chul Yoo, Virginia Urquidi, David M. Lubman, Steve Goodison

Research output: Contribution to journalArticle

47 Scopus citations

Abstract

A combination of LC and MS was applied to an isogenic breast tumor metastasis model to identify proteins associated with a cellular phenotype. Chromatofocusing followed by nonporous-RP-HPLC/ESI-TOF MS was applied to cell lysates of a pair of monoclonal cell lines from the human breast carcinoma cell line MDA-MB-435 that have different metastatic phenotypes in immune-compromised mice. This method was developed to separate proteins based on pI and hydrophobicity. The high resolution and mass accuracy of ESI-TOF measurements provided a good correlation of theoretical MW and experimental Mr values of intact proteins measured in mass maps obtained in the pH range 3.8-6.4. The isolated proteins were digested by trypsin and analyzed by MALDI-TOF MS, MALDI-QIT-TOF MS, and monolith-based HPLC/MS/MS. The unique combination of the techniques provided valuable information including quantitation and modification of proteins. We identified 89 selected proteins, of which 43 were confirmed as differentially expressed. Metastasis-associated proteins included galectin-1, whereas annexin I and annexin II were associated with the nonmetastatic phenotype. In this study, we demonstrate that combining a variety of MS tools with a multidimensional liquid-phase separation provides the ability to map cellular protein content, to search for modified proteins, and to correlate protein expression with cellular phenotype.

Original languageEnglish (US)
Pages (from-to)299-312
Number of pages14
JournalProteomics
Volume7
Issue number2
DOIs
StatePublished - Jan 1 2007

Keywords

  • Cancer biomarkers
  • Electrospray ionization time-of-flight mass spectrometry
  • Liquid chromatography
  • Metastasis
  • Monolith

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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