Two-dimensional gel electrophoresis maps of the proteome and phosphoproteome of primitively cultured rat mesangial cells

被引:25
作者
Jiang, XS [1 ]
Tang, LY [1 ]
Cao, XJ [1 ]
Zhou, H [1 ]
Xia, QC [1 ]
Wu, JR [1 ]
Zeng, R [1 ]
机构
[1] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Res Ctr Proteome Anal,Key Lab Proteom,Grad Sch, Shanghai 200031, Peoples R China
关键词
mass spectrometry; mesangial cell; phosphoproteome; proteome; two-dimensional electrophoresis;
D O I
10.1002/elps.200500286
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Mesangial cells (MC) play an important role in maintaining the structure and function of the glomerulus. The proliferation of MC is a prominent feature of many kinds of glomerular disease. The first reference 2-DE maps of rat mesangial cells (RMC), stained with silver staining or Pro-Q Diamond dye, have been established here to describe the proteome and phosphoproteome of RMC, respectively. A total of 157 selected protein spots, corresponding to 118 unique proteins, have been identified by MALDI-TOF-MS or LC-ESI-IT-MS/MS, in which 37 protein spots representing 28 unique proteins have also been stained with Pro-Q Diamond, indicating that they are in phosphorylated forms. All the identified proteins were bioinformatically annotated in detail according to their physiochemical characteristics, subcellular location, and function. Most of the separated or identified protein spots are distributed in the area of mass 10-70 kDa and p/5.0-8.0. The identified proteins include mainly cytoplasmic and nuclear proteins and some mitochondrial, endoplasmic reticulum, and membrane proteins. These proteins are classified into different functional groups such as structure and mobility proteins (21.2%), metabolic enzymes (16.9%), protein folding and metabolism proteins (13.6%), signaling proteins (14.4%), heat-shock proteins (7.6%), and other functional proteins (12.7%). While structure and mobility proteins are mostly represented by protein spots with high abundance, signaling proteins are mostly represented by protein spots with relatively low abundance. Such a 2-DE database for RMC, especially with many signaling proteins and phosphoproteins characterized, will provide a valuable resource for comparative proteomics analysis of normal and pathologic conditions affecting MC function or pathologic progress.
引用
收藏
页码:4540 / 4562
页数:23
相关论文
共 48 条
[21]   LPA as a determinant of mesangial growth and apoptosis [J].
Inoue, CN .
SEMINARS IN NEPHROLOGY, 2002, 22 (05) :415-422
[22]   A comparative proteomic strategy for subcellular proteome research - ICAT approach coupled with bioinformatics prediction to ascertain rat liver mitochondrial proteins and indication of mitochondrial localization for catalase [J].
Jiang, XS ;
Dai, J ;
Sheng, QH ;
Zhang, L ;
Xia, QC ;
Wu, JR ;
Zeng, R .
MOLECULAR & CELLULAR PROTEOMICS, 2005, 4 (01) :12-34
[23]   A high-throughput approach for subcellular proteome - Identification of rat liver proteins using subcellular fractionation coupled with two-dimensional liquid chromatography tandem mass spectrometry and bioinformatic analysis [J].
Jiang, XS ;
Zhou, H ;
Zhang, L ;
Sheng, QH ;
Li, SJ ;
Li, L ;
Hao, P ;
Li, YX ;
Xia, QC ;
Wu, JR ;
Zeng, R .
MOLECULAR & CELLULAR PROTEOMICS, 2004, 3 (05) :441-455
[24]   Protein kinase A-catalyzed phosphorylation of heat shock protein 60 chaperone regulates its attachment to histone 2B in the T lymphocyte plasma membrane [J].
Khan, IU ;
Wallin, R ;
Gupta, RS ;
Kammer, GM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (18) :10425-10430
[25]  
KRIZ W, 1995, J AM SOC NEPHROL, V5, P1731
[26]   Predicting transmembrane protein topology with a hidden Markov model: Application to complete genomes [J].
Krogh, A ;
Larsson, B ;
von Heijne, G ;
Sonnhammer, ELL .
JOURNAL OF MOLECULAR BIOLOGY, 2001, 305 (03) :567-580
[27]   Mesangial cell proliferation inhibitors for the treatment of proliferative glomerular disease [J].
Kurogi, Y .
MEDICINAL RESEARCH REVIEWS, 2003, 23 (01) :15-31
[28]   A SIMPLE METHOD FOR DISPLAYING THE HYDROPATHIC CHARACTER OF A PROTEIN [J].
KYTE, J ;
DOOLITTLE, RF .
JOURNAL OF MOLECULAR BIOLOGY, 1982, 157 (01) :105-132
[29]   Study of human laryngeal muscle protein using two-dimensional electrophoresis and mass spectrometry [J].
Li, ZB ;
Lehar, M ;
Braga, N ;
Westra, W ;
Liu, LH ;
Flint, PW .
PROTEOMICS, 2003, 3 (07) :1325-1334
[30]   Heat-shock protein-25/27 phosphorylation by the δ isoform of protein kinase C [J].
Maizels, ET ;
Peters, CA ;
Kline, M ;
Cutler, RE ;
Shanmugam, M ;
Hunzicker-Dunn, M .
BIOCHEMICAL JOURNAL, 1998, 332 :703-712