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中国生物工程杂志

China Biotechnology
China Biotechnology  2010, Vol. 30 Issue (02): 27-31    DOI:
    
Establishment of Proteomics Methods of Golgi Complex Isolated from Gastric Cancer Cells
HE Ting-ting,YI Yong-fen,LI Yan-qing,XIAO Zhong
Molecular Medicine and Cancer Research,Department of Pathology, Chongqing Medical University, Chongqing 400016, China
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Abstract  

The proteomics technique platform of Golgi apparatus isolated from gastric cancer cells was established successfully by using subcellular isolation with sucrose density gradient ultra-centrifugation, the separation of proteins of Golgi complex by 2-DE, the analysis of Electrophoresis patterns with ImageMaster 2D software, and the identification of proteins based on matrix-assisted laser desorption/ionization time of flight mass spectrometry. The results indicated that the well-resolved and reproducible 2-DE pattern of Golgi apparatus isolated from gastric cancer cells was obtained , and twelve proteins were identified, including proteins related to protein synthesis and folding, membrane fusion proteins, regulatory proteins, transportation proteins, proteins related to apoptosis, proteins related to cell proliferation. The first installment of the proteome technology of Golgi complex isolated from gastric cancer cells SGC7901 by 2-DE and MALDI-TOF MS will benefit for further investigation on the functions of Golgi complex.



Key wordsGastric cancer cell      Golgi apparatus      Subcellular proteome      Subcellular separation     
Received: 13 November 2009      Published: 26 February 2010
Cite this article:

HE Ting-Ting, YI Yong-Fen, LI Yan-Jing, XIAO Zhong. Establishment of Proteomics Methods of Golgi Complex Isolated from Gastric Cancer Cells. China Biotechnology, 2010, 30(02): 27-31.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2010/V30/I02/27

[1] Rabouille C, Jokitalo E. Golgi apparatus partitioning during cell division. Mol Memb Biol,2003,20(2):117127. 
[2] Mukherjee S, Chiu R, Leung S M, et al. Fragmentation of the Golgi apparatus:An early apoptotic event independent of the cytoskeleton. Traffic,2007,8(4): 369–378. 
[3] Taylor R S, Fialka I, Jones S M, et al. Twodimensional mapping of the endogenous proteins of the rat hepatocyte Golgi complex cleared of proteins in transit. Electrophoresis,1997,18(14): 26012612. 
[4] Gaston D, Tsaousis A D, Roger AJ. Predicting proteomes of mitochondria and related organelles from genomic and expressed sequence tag data. Methods Enzymol, 2009,457:2147. 
[5] 黄培堂.细胞实验指南.北京: 科学出版社, 2003.356370. Huang P T.Manual.Baijing:Science Press,2003.350~370. 
[6] Balch W E, Dunphy W G, Braell WA, et al. Reconstitution of the transport of protein between successive compartments of the golgi measured by the coupled incorporation of nacetylglucosamine. Cell,1984,39 (2 Pt 1): 405416. 
[7] Venky S R, Arjun K S, Ram R. The Saccharomyces cerevisiae PHM8 gene encodes a soluble magnesiumdependent lysophosphatidic acid phosphatase. J Biol Chem, 2008, 283: 8846  8854. 
[8] Graham J M, Higgins J A. Biomembrane protocols Ⅰ. Isolation and analysis.In:Methods in Molecular Biology.Totowa: Humana Press Inc,1993.19:100102. 
[9] Yan J X, Wait R, Berkelman T, et al. A modified silver staining protocol for visualization of proteins compatible with matricassisted laser desorption/ionization and electrospray ionization mass spectrometry. Electrophoresis,2000,21(17):36663672. 
[10] lowenthal M S, howell K E, Wu C C. Proteomic analysis of the stacked Golgi complex. Methods Mol Biol,2008,432:3749. 
[11] Wu C C, Yates J R, Neville M C, et al. Proteomic analysis of two functional states of Golgi complex in mammary egithelial cells. Traffic,2000,1:769782. 
[12]Wu C C, MacCoss M J, Mardones G, et al. Organellar proteomics reveals Golgi arginine dimethylation. Mol Biol Cell, 2004, 15(6): 29072919. 
[13] Takatalo M S, Kouvonen P, Corthals S G, et al. Identification of new Golgi complex specific proteins by direct organelle proteomic analysis. Proteomics,2006,6: 35023508. 
[14] Brunner Y, Schvartz D, Couté Y, et al. Proteomics of regulated secretory organelles. Mass Spectrom Rev,2009,28(5):844867. 
[15] Michelsen U, von Hagen J. Isolation of subcellular organelles and structures. Methods Enzymol,2009,463:305328. 
[16] Zaks W J, Creutz C E. Evaluation of the annexin as potential mediators of membrane fusion in exocytosis. J Bioenerg Biomembr,1990,22:97120. 
[17] Das S, Sierra J C, Soman K V, et a1.Diffrentia1 protein expression profiles of gastric epithelia1 cells following Helicobacter pylori infection using protein chips.J Proteome Res,2005,3:920930. 
[18] Emoto K, Sawada H, Yamada Y, et al. Annexin II overexpression is correlated with poor prognosis in human gastric carcinoma. Anticancer Res,2001,21(2B):13391345. 
[19] Chan C H, Ko C C, Chang J G, et al. Subcellular and functionalproteomic analysis of the cellular responses induced by Helicobacter pylori. Mol Cell Proteomics,2006,5: 702713.

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