Identification of Fibrosis-Relevant Proteins Using DIGE (Difference in Gel Electrophoresis) in Different Models of Hepatic Fibrosis

Abstract

Proteomics became a more and more important technique for the large-scale analysis of proteins during the last years. Two-dimensional (2D) electrophoresis as a major tool of proteomics is a powerful method to compare two different biological stages (e. g. healthy and diseased tissue) and to find differences in their protein pattern. One major problem in proteomics is the gel to gel variation of two-dimensional gel electrophoresis, which could cause artefacts in the detection of expression differences. The "difference in gel electrophoresis" (DIGE) technique allows the separation of two proteomes in the same gel. The protein pools were labelled with different fluorescent dyes and equal amounts of protein were separated in the same gel. Another advantage of DIGE is the possibility to separate an internal standard labelled with a third dye in the same gel to allow quantitative expression analysis. We compared proteomes of three different fibrosis models with the appropriate control (tissue inhibitor of metalloproteinases-1 (TIMP-1) overexpressing HepG2 cells in comparison to a HepG2 control, freshly isolated HSC in comparison to activated HSC and healthy mouse liver in comparison to fibrotic mouse liver). Among the differentially expressed proteins several were already found to be relevant for fibrosis but we also detected some proteins like the selenium binding protein 2 which might be relevant for hepatic fibrosis.

A Proteomics Study of Brassinosteroid Response in Arabidopsis

“…The protein identities of spots of interest are subsequently determined by mass spectrometry. 2-D DIGE has been used widely in quantitative proteomics studies in a wide range of biological and disease systems (35)(36)(37)(38)(39)(40)(41)(42). In plants, 2-D DIGE has been used to identify proteins regulated by UV-B light (43), salt and osmotic stresses (44), cold stress (45), fungal elicitors (46), and gibberellins (47).…”

Proteomic Profiling of Differentially Expressed Proteins from Bax inhibitor-1 Knockout and Wild Type Mice

“…SBP2 levels decreased dramatically in livers of C57BL/6J mice with peroxisome proliferator-activated receptorbinding properties for selenium, and SBP2 is mainly detectable in liver cell-specific pleiotropic responses, including the development of liver tumors (Chu et al, 2004). SBP2 is also relevant for hepatic fibrosis (Henkel et al, 2005) and has been identified as a major hepatic target for acetaminophen, while demonstrating gender differences in protein abundance (Mattow et al, 2006). In bi-1 -/-mice, SBP2 expression was relatively higher compared to bi-1 +/+ mice.…”

Sex-dependent compensated oxidative stress in the mouse liver upon deletion of catechol O-methyltransferase

“…PPARα activation is important for the induction of cell specific pleiotropic responses, including liver tumours [65]. In addition, SBP2 may play a role in fibrosis [66]. Despite these findings, the precise biological function of SBP2 is scarcely known and its role in catecholamine metabolism is unidentified.…”