蛋白质组学研究方法及应用的研究进展

摘要/Abstract

摘要： 蛋白质是细胞生理功能的具体执行者，并常以蛋白复合体或与核酸相互作用的形式来发挥作用。蛋白质组代表在一定时间和特定条件下，1个细胞、组织或生物个体所表达的全部蛋白，因此蛋白质组会随着时间和空间的改变而发生动态变化。蛋白质组学从整体水平研究蛋白质组的结构、功能、表达模式及其相互作用的方式。根据研究内容的不同，蛋白质组学可分为表达蛋白质组学、结构蛋白质组学和功能蛋白质组学等。研究蛋白质组学有助于了解蛋白的结构、细胞的功能、生命的本质及活动规律，为疾病的诊断、治疗、疫苗及新药开发提供科学依据。研究蛋白质组学的常用手段主要有：双向聚丙烯酰胺凝胶电泳、质谱、酵母双杂交系统及蛋白芯片法等，文章将对蛋白质组学常用的技术方法及应用进行综述。

关键词: 蛋白质组学; 双向凝胶电泳; 质谱; 蛋白质分析

Abstract: Proteins are the chief performers for physiological functions in a cell, and often function as protein complexes or interact with nucleic acids. A proteome represents all the proteins expressed by a cell, a tissue, or an organism at a defined time point under certain circumstances. Therefore, proteomes will undergo dynamic changes with alterations in time and space. The aim of proteomics is to globally study structures, functions and expression modes of proteome as well as interactions among proteins in a proteome. Based on the contents of study, proteomics can be classified as expression proteomics, structural proteomics and functional proteomics,respectively. The study of proteomes will help understand structures of proteins, functions of cells, nature and active rules of life, and will provide scientific basis for diagnosis and treatment of diseases as well as development of novel drugs. The techniques commonly used in proteomic studies include two-dimentional polyacrylamide gel electrophoresis, mass spectrometry, yeast two-hybrid system and protein chips. This article will provide a brief overview of proteomics and techniques frequently utilized for proteomic studies.

Key words: proteomics; two-dimentional gel electrophoresis; mass spectrometry; protein analysis

中图分类号:

谭伟, 黄莉, 谢芝勋. 蛋白质组学研究方法及应用的研究进展[J]. 中国畜牧兽医, 2014, 41(9): 40-46.

TAN Wei, HUANG Li, XIE Zhi-xun. Research Progress on Proteomic Techniques and their Applications[J]. , 2014, 41(9): 40-46.

参考文献

1 孙成友,李娟,刘雨田,等. 蛋白质组学技术在病毒及其感染机制研究中的应用[J]. 中国畜牧兽医,2013,40(5):75～79.

2 张太翔,田国宁,张金玲,等. 蛋白质组学与神经系统疾病的研究进展[J]. 中国畜牧兽医,2008,35(7):46～48.

3 党源. 新城疫病毒单感染及猪流感病毒与猪链球菌共感染差异蛋白质组学分析[D]. 长春:吉林大学,2013.

4 高昭辉,董书伟,薛慧文,等. 纳米铜对大鼠肝脏毒性的蛋白质组2-DE图谱分析[J]. 中国畜牧兽医,2012,39(11):22～26.

5 Abel L, Kutschki S, Turewicz M, et al. Autoimmune profiling with protein microarrays in clinical applications[J]. Biochim Biophys Acta, 2014, 1844(5): 977～987.

6 Altelaar A F, Munoz J, Heck A J. Next-generation proteomics: Towards an integrative view of proteome dynamics[J]. Nat Rev Genet, 2013, 14(1): 35～48.

7 Alvarez-Chaver P, Otero-Estévez O, Páez de la Cadena M, et al. Proteomics for discovery of candidate colorectal cancer biomarkers[J]. World J Gastroenterol, 2014, 20(14): 3804～3824.

8 Andersen J S, Mann M. Functional genomics by mass spectrometry[J]. FEBS Lett, 2000, 480(1): 25～31.

9 Anderson L, Seilhamer J. A comparison of selected mRNA and protein abundances in human liver[J]. Electrophoresis, 1997, 18(3～4): 533～537.

10 Arenkov P, Kukhtin A, Gemmell A, et al. Protein microchips: Use for immunoassay and enzymatic reactions[J]. Anal Biochem, 2000, 278(2): 123～131.

11 Banks R E, Dunn M J, Hochstrasser D F, et al. Proteomics: New perspectives, new biomedical opportunities[J]. Lancet, 2000, 356(9243): 1749～1756.

12 Beck M, Claassen M, Aebersold R. Comprehensive proteomics[J]. Curr Opin Biotechnol, 2011, 22(1): 3～8.

13 Brewis I A, Brennan P. Proteomics technologies for the global identification and quantification of proteins[J]. Adv Protein Chem Struct Biol, 2010, 80: 1～44.

14 Cho A, Normile D. Nobel prize in chemistry. Mastering macromolecules[J]. Science, 2002, 298(5593): 527～528.

15 Cho W C. Proteomics technologies and challenges[J]. Genomics Proteomics Bioinformatics, 2007, 5(2): 77～85.

16 de Hoog C L, Mann M. Proteomics[J]. Annu Rev Genomics Hum Genet, 2004, 5: 267～293.

17 Ding S J, Qian W J, Smith R D. Quantitative proteomic approaches for studying phosphotyrosine signaling[J]. Expert Rev Proteomics, 2007, 4(1): 13～23.

18 Domon B, Aebersold R. Mass spectrometry and protein analysis[J]. Science, 2006, 312(5771): 212～217.

19 Fenn J B, Mann M, Meng C K, et al. Electrospray ionization for mass spectrometry of large biomolecules[J]. Science,1989, 246(4926): 64～71.

20 Fields S, Song O. A novel genetic system to detect protein-protein interactions[J]. Nature, 1989, 340(6230): 245～246.

21 Godovac-Zimmermann J, Brown L R. Perspectives for mass spectrometry and functional proteomics[J]. Mass Spectrom Rev, 2001, 20(1): 1～57.

22 Graves P R, Haystead T A. Molecular biologist's guide to proteomics[J]. Microbiol Mol Biol Rev, 2002, 66(1): 39～63.

23 Gygi S P, Rochon Y, Franza B R, et al. Correlation between protein and mRNA abundance in yeast[J]. Mol Cell Biol, 1999, 19(3): 1720～1730.

24 Gygi S P, Corthals G L, Zhang Y, et al. Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology[J]. Proc Natl Acad Sci USA, 2000, 97(17): 9390～9395.

25 Hanash S. Disease proteomics[J]. Nature, 2003, 422(6928): 226～232.

26 Hendrickson R C, Douglass J F, Reynolds L D, et al. Mass spectrometric identification of mtb81, a novel serological marker for tuberculosis[J]. J Clin Microbiol, 2000, 38(6): 2354～2361.

27 Karas M, Hillenkamp F. Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons[J]. Anal Chem, 1988, 60(20): 2299～2301.

28 Klose J. Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals[J]. Humangenetik, 1975, 26(3): 231～243.

29 Kopf E, Zharhary D. Antibody arrays——An emerging tool in cancer proteomics[J]. Int J Biochem Cell Biol, 2007, 39(7～8): 1305～1317.

30 Krishna R G, Wold F. Post-translational modification of proteins[J]. Adv Enzymol Relat Areas Mol Biol, 1993, 67: 265～298.

31 MacBeath G, Schreiber S L. Printing proteins as microarrays for high-throughput function determination[J]. Science, 2000, 289(5485): 1760～1763.

32 Maxwell K L, Frappier L. Viral proteomics[J]. Microbiol Mol Biol Rev, 2007, 71(2): 398～411.

33 May C, Brosseron F, Chartowski P, et al. Instruments and methods in proteomics[J]. Methods Mol Biol, 2011, 696: 3～26.

34 McKerrow J H, Bhargava V, Hansell E, et al. A functional proteomics screen of proteases in colorectal carcinoma[J]. Mol Med, 2000, 6(5): 450～460.

35 Nishimura T, Ogiwara A, Fujii K, et al. Disease proteomics toward bedside reality[J]. J Gastroenterol, 2005, 40(Suppl)16: 7～13.

36 O'Donovan C, Apweiler R, Bairoch A. The human proteomics initiative (HPI)[J]. Trends Biotechnol, 2001, 19(5): 178～181.

37 O'Farrell P H. High resolution two-dimensional electrophoresis of proteins[J]. J Biol Chem, 1975, 250(10): 4007～4021.

38 Patton W F. Detection technologies in proteome analysis[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2002, 771(1～2): 3～31.

39 Paweletz C P, Charboneau L, Bichsel V E, et al. Reverse phase protein microarrays which capture disease progression show activation of pro-survival pathways at the cancer invasion front[J]. Oncogene, 2001, 20(16): 1981～1989.

40 Petschnigg J, Snider J, Stagljar I. Interactive proteomics research technologies: Recent applications and advances[J]. Curr Opin Biotechnol, 2011, 22(1): 50～58.

41 Pratsch K, Wellhausen R, Seitz H. Advances in the quantification of protein microarrays[J]. Curr Opin Chem Biol, 2014, 18: 16～20.

42 Rao V S, Srinivas K, Sujini G N, et al. Protein-protein interaction detection: Methods and analysis[J]. Int J Proteomics, 2014, 2014: 147648.

43 Rappsilber J, Mann M. What does it mean to identify a protein in proteomics[J]. Trends Biochem Sci, 2002, 27(2): 74～78.

44 Robinson W H, DiGennaro C, Hueber W, et al. Autoantigen microarrays for multiplex characterization of autoantibody responses[J]. Nat Med, 2002, 8(3): 295～301.

45 Rodrigo M A, Zitka O, Krizkova S, et al. MALDI-TOF MS as evolving cancer diagnostic tool: A review[J]. J Pharm Biomed Anal, 2014, 95: 245～255.

46 Scheele G A. Two-dimensional gel analysis of soluble proteins. Charaterization of guinea pig exocrine pancreatic proteins[J]. J Biol Chem, 1975, 250(14): 5375～5385.

47 Snider J, Kittanakom S, Damjanovic D, et al. Detecting interactions with membrane proteins using a membrane two-hybrid assay in yeast[J]. Nat Protoc, 2010, 5(7): 1281～1293.

48 Thongboonkerd V. Proteomics[J]. Forum Nutr, 2007, 60: 80～90.

49 Uetz P, Giot L, Cagney G, et al. A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae[J]. Nature, 2000, 403(6770): 623～627.

50 Unlü M, Morgan M E, Minden J S. Difference gel electrophoresis: A single gel method for detecting changes in protein extracts[J]. Electrophoresis, 1997, 18(11): 2071～2077.

51 Wasinger V C, Cordwell S J, Cerpa-Poljak A, et al. Progress with gene-product mapping of the Mollicutes: Mycoplasma genitalium[J]. Electrophoresis, 1995, 16(7): 1090～1094.

52 Weinrich D, Jonkheijm P, Niemeyer C M, et al. Applications of protein biochips in biomedical and biotechnological research[J]. Angew Chem Int Ed Engl, 2009, 48(42): 7744～7751.

53 Wilkins M R, Sanchez J C, Gooley A A, et al. Progress with proteome projects: Why all proteins expressed by a genome should be identified and how to do it[J]. Biotechnol Genet Eng Rev, 1996, 13: 19～50.

54 Xiao Z, Jiang X, Beckett M L, et al. Generation of a baculovirus recombinant prostate-specific membrane antigen and its use in the development of a novel protein biochip quantitative immunoassay[J]. Protein Expr Purif, 2000, 19(1): 12～21.

55 Zhu H, Bilgin M, Bangham R, et al. Global analysis of protein activities using proteome chips[J]. Science, 2001, 293(5537): 2101～2105.

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