The functions of the liver and the pancreas differ; however, chronic inflammation in both organs is associated with fibrosis. Evidence suggests that fibrosis in both organs is partially regulated by organ-specific stellate cells. We explore the proteome of human hepatic stellate cells (hHSC) and human pancreatic stellate cells (hPaSC) using mass spectrometry (MS)-based quantitative proteomics to investigate pathophysiologic mechanisms. Proteins were isolated from whole cell lysates of immortalized hHSC and hPaSC. These proteins were tryptically digested, labeled with tandem mass tags (TMT), fractionated by OFFGEL, and subjected to MS. Proteins significantly different in abundance (P<0.05) were classified via gene ontology (GO) analysis. We identified 1223 proteins and among them, 1222 proteins were quantifiable. Statistical analysis determined that 177 proteins were of higher abundance in hHSC, while 157 were of higher abundance in hPaSC. GO classification revealed that proteins of relatively higher abundance in hHSC were associated with protein production, while those of relatively higher abundance in hPaSC were involved in cell structure. Future studies using the methodologies established herein, but with further upstream fractionation and/or use of enhanced MS instrumentation will allow greater proteome coverage, achieving a comprehensive proteomic analysis of hHSC and hPaSC.
|Number of pages||9|
|Journal||Genomics, Proteomics and Bioinformatics|
|State||Published - Apr 2013|
Bibliographical noteFunding Information:
Funds were provided by the following NIH grants: 1 F32 DK085835-01A1 (JP), 1 R21 DK081703-01A2 (DC), 5 P30 DK034854-24 (Harvard Digestive Diseases Center; DC), as well as a grant from the American College of Gastroenterology: ACG – 042103580 (JP). We would like to thank the Burrill family for their generous support through the Burrill Research Grant. We would also like to thank members of the Steen Laboratory at Boston Children’s Hospital, in particular John FK Sauld and Ali Ghoulidi for their technical assistance and critical reading of the manuscript. In addition, we thank members of the Center for Pancreatic Disease at Brigham and Women’s Hospital, particularly Shadeah Suleiman for her technical assistance.
- Tandem mass tag
ASJC Scopus subject areas
- Molecular Biology
- Computational Mathematics