Proteomic analysis of a rat pancreatic stellate cell line using liquid chromatography tandem mass spectrometry (LC-MS/MS)

Joao A. Paulo, Raul Urrutia, Peter A. Banks, Darwin L. Conwell, Hanno Steen

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Pancreatic stellate cells (PaSC) are emerging as key mediators in chronic pancreatitis and pancreatic cancer pathogenesis. Proteins regulating the biomolecular pathways involved in the conversion of quiescent to activated PaSC may have a significant influence on the development of chronic pancreatitis. We aim to compare differentially expressed proteins in activated and serum-starved non-proliferating PaSC using a mass spectrometry-based proteomics strategy. We cultured an immortalized rat PaSC cell line in media supplemented with 10% fetal bovine serum and in serum-free media. Using gel-based mass spectrometry (GeLC-MS/MS), we identified nearly 1500 proteins. Qualitative and quantitative proteomic analysis revealed several hundred proteins as differentially abundant between the two cell states. Proteins of greater abundance in activated PaSC included isoforms of actin (e.g., smooth muscle actin) and ribosomal proteins. Conversely, proteins more abundant in non-proliferating PaSC than in activated PaSC included signaling proteins MAP kinase 3 and Ras-related proteins. In addition, we have determined the molecular functions and biological pathways for these proteins. We are confident that the application of mass spectrometry-based strategies, such as that described herein, to investigate specific proteins in PaSC may lead to a better understanding of the molecular mechanisms involved in pancreatic diseases, such as chronic pancreatitis.

Original languageEnglish (US)
Pages (from-to)708-717
Number of pages10
JournalJournal of Proteomics
Volume75
Issue number2
DOIs
StatePublished - Dec 21 2011

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Pancreatic Stellate Cells
Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Proteomics
Mass spectrometry
Rats
Cells
Cell Line
Chronic Pancreatitis
Proteins
Mass Spectrometry
Actins
ras Proteins
Pancreatic Diseases
Muscle Proteins
Ribosomal Proteins
Serum-Free Culture Media
Cell signaling
Serum

Keywords

  • Biomarker
  • Fibrosis
  • Pancreatitis
  • Proteomics
  • Stellate cells

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Proteomic analysis of a rat pancreatic stellate cell line using liquid chromatography tandem mass spectrometry (LC-MS/MS). / Paulo, Joao A.; Urrutia, Raul; Banks, Peter A.; Conwell, Darwin L.; Steen, Hanno.

In: Journal of Proteomics, Vol. 75, No. 2, 21.12.2011, p. 708-717.

Research output: Contribution to journalArticle

Paulo, Joao A. ; Urrutia, Raul ; Banks, Peter A. ; Conwell, Darwin L. ; Steen, Hanno. / Proteomic analysis of a rat pancreatic stellate cell line using liquid chromatography tandem mass spectrometry (LC-MS/MS). In: Journal of Proteomics. 2011 ; Vol. 75, No. 2. pp. 708-717.
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