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

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

Research output: Contribution to journalArticle

47 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Proteomics
Tumors
Breast Neoplasms
Neoplasm Metastasis
Proteins
Phenotype
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
High Pressure Liquid Chromatography
Cells
Annexin A2
Galectin 1
Annexin A1
Cell Line
Hydrophobicity
Hydrophobic and Hydrophilic Interactions
Phase separation
Trypsin
Liquids

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Proteomic profiling identifies breast tumor metastasis-associated factors in an isogenic model. / Kreunin, Paweena; Yoo, Chul; Urquidi, Virginia; Lubman, David M.; Goodison, Steven.

In: Proteomics, Vol. 7, No. 2, 01.01.2007, p. 299-312.

Research output: Contribution to journalArticle

Kreunin, Paweena ; Yoo, Chul ; Urquidi, Virginia ; Lubman, David M. ; Goodison, Steven. / Proteomic profiling identifies breast tumor metastasis-associated factors in an isogenic model. In: Proteomics. 2007 ; Vol. 7, No. 2. pp. 299-312.
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