柴政斌;张更林;韩金祥;

• 1:卫生部生物技术药物重点实验室山东省罕少见病重点实验室山东省医药生物技术研究中心山东省医学科学院
• 2:济南大学山东省医学科学院医学与生命科学学院

摘要(Abstract)：

蛋白间相互作用在细胞的生命活动中起着关键作用,GST-pull down技术被广泛用于体外验证蛋白间的直接相互作用。GST-pull down技术既可用来验证已知蛋白间的相互作用,也可用来寻找与已知蛋白相互作用的未知蛋白。虽然该技术在体外验证蛋白间的相互作用上有着较强的特异性,但下结论时仍需慎重。本文就GST-pull down技术的原理及其应用及该技术在应用中需要注意的部分问题进行综述。

关键词(KeyWords)： GST-pull down;蛋白质相互作用;融合蛋白

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 柴政斌;张更林;韩金祥;

DOI: 10.13200/j.cnki.cjb.000628

参考文献(References)：

• [1]Wissmueller S,Font J,Liew CW,et al.Protein-protein interactions:analysis of a false positive GST pulldown result[J].Proteins,2011,79(8):2365-2371.
• [2]Smith DB,Johnson KS.Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione Stransferase[J].Gene,1988,67(1):31-40.
• [3]Chen AJ,Wang S,Zhao XF,et al.Enzyme E2 from Chinese white shrimp inhibits replication of white spot syndrome virus and ubiquitinates its RING domain proteins[J].J Virol,2011,85(16):8069-8079.
• [4]Liu PC,Wang JX,Song QS,et al.The participation of calponin in the cross talk between 20-hydroxyecdysone and juvenile hormone signaling pathways by phosphorylation variation[J].PLoS One,2011,6(5):e19776.
• [5]Harper S,Speicher DW.Expression and purification of GST fusipn proteins//Coligan JE,Dunn BM,Speicher DW,et al.Current protocols in protein science.New York:John Wiley&Sons,Inc.,1997:6.6.1-6.6.21.
• [6]Hong JP,Byun MY,An K,et al.OsKu70 is associated with developmental growth and genome stability in rice[J].Plant Physiol,2010,152(1):374-387.
• [7]Maekawa S,Kobayashi Y,Odagaki S,et al.Interaction of NAP-22 with brain glutamic acid decarboxylase(GAD)[J].Neurosci Lett,2013,537(1):50-54.
• [8]Zhang Y,Wang L.Characterization of the mitochondrial localization of the nuclear receptor SHP and regulation of its subcellular distribution by interaction with Bcl2 and HNF4alpha[J].PLoS One,2013,8(7):e68491.
• [9]Banko MI,Krzyzanowski MK,Turcza P,et al.Identification of amino acid residues of ERH required for its recruitment to nuclear speckles and replication foci in HeLa cells[J].PLoS One,2013,8(8):e74885.
• [10]Bornert O,M尴ller TC,Boeuf J,et al.Identification of a novel protein-protein interaction motif mediating interaction of GPCRassociated sorting proteins with G protein-coupled receptors[J].PLoS One,2013,8(2):e56336.
• [11]Shen SL,Qiu FH,Dayarathna TK,et al.Identification of Dermcidin as a novel binding protein of Nck1 and characterization of its role in promoting cell migration[J].Biochim Biophys Acta,2011,1812(6):703-710.
• [12]Zhang T,Xie N,He W,et al.An integrated proteomics and bioinformatics analyses of hepatitis B virus X interacting proteins and identification of a novel interactor apoA-I[J].J Proteomics,2013,84(1):92-105.
• [13]Jones N,Dumont DJ.The Tek/Tie2 receptor signals through a novel Dok-related docking protein,Dok-R[J].Oncogene,1998,17(9):1097-1108.
• [14]Trinkle-Mulcahy L.Resolving protein interactions and complexes by affinity purification followed by label-based quantitative mass spectrometry[J].Proteomics,2012,12(10):1623-1638.
• [15]Borth N,Litsche K,Franke C,et al.Functional interaction between type III-secreted protein IncA of Chlamydophila psittaci and human G3BP1[J].PLoS One,2011,6(1):e16692.
• [16]Einarson MB,Pugacheva EN,Orlinick JR.Identification of protein-protein interactions with glutathione-S-transferase(GST)fusion proteins[J].CSH Protoc,2007,2007:pdb top11.
• [17]Fletcher S,Bowden SE,Marrion NV.False interaction of syntaxin 1A with a Ca(2+)-activated K(+)channel revealed by coimmunoprecipitation and pull-down assays:implications for identification of protein-protein interactions[J].Neuropharmacology,2003,44(6):817-827.
• [18]Mackay JP,Sunde M,Lowry JA,et al.Protein interactions:is seeing believing[J].Trends Biochem Sci,2007,32(12):530-531.
• [19]Vikis HG,Guan KL.Glutathione-S-transferase-fusion based assays for studying protein-protein interactions[J].Methods Mol Biol,2004,261(1):175-186.
• [20]Harper S,Speicher DW.Purification of proteins fused to glutathione S-transferase[J].Methods Mol Biol,2011,681(1):259-280.
• [21]Gong Z,Zhuo Q,Wu J,et al.Study on no-fusion expression and purification of glycine max dehydration-responsive elementbinding protein(GmDREB1)expressed in E.coli[J].Wei Sheng Yan Jiu,2013,42(3):392-398.
• [22]Huang X,Wang J,Xia W,et al.Prokaryotic expression,purification and functional characterization of human FHL3[J].Biotechnol Lett,2009,31(10):1499-1504.
• [23]Donovan RS,Robinson CW,Glick BR.Optimizing the expression of a monoclonal antibody fragment under the transcriptional control of the Escherichia coli lac promoter[J].Can J Microbiol,2000,46(6):532-541.
• [24]VolontèF,Marinelli F,Gastaldo L,et al.Optimization of glutaryl-7-aminocephalosporanic acid acylase expression in E.coli[J].Protein Expr Purif,2008,61(2):131-137.
• [25]Jin J,Fan M,Li Y.Optimizing expression and antibody preparation of recombinant Streptococcus mutans surface protein[J].Hua Xi Kou Qiang Yi Xue Za Zhi,2012,30(3):251-254,258.
• [26]Sadraeian M,Ghoshoon MB,Mohkam M,et al.Modification in media composition to obtain secretory production of STxBbased vaccines using Escherichia coli[J].Virol Sin,2013,28(1):43-48.
• [27]Kim MJ,Park HS,Seo KH,et al.Complete solubilization and purification of recombinant human growth hormone produced in Escherichia coli[J].PLoS One,2013,8(2):e56168.
• [28]Du C,Han L,Xie Z.Cloning,expression in Escherichia coli,and purification of soluble recombinant duck interleukin-2[J].World J Microbiol Biotechnol,2012,28(4):1495-1501.
• [29]Flick K,Ahuja S,Chene A,et al.Optimized expression of Plasmodium falciparum erythrocyte membrane protein 1 domains in Escherichia coli[J].Malar J,2004,3(1):50-57.
• [30]Wang Z,Xiang Q,Wang G,et al.Optimizing expression and purification of an ATP-binding gene gsiA from Escherichia coli k-12 by using GFP fusion[J].Genet Mol Biol,2011,34(4):661-668.
• [31]Gauci C,Jenkins D,Lightowlers MW.Strategies for optimal expression of vaccine antigens from Taeniid cestode parasites in Escherichia coli[J].Mol Biotechnol,2011,48(3):277-289.
• [32]Liew CK,Gamsjaeger R,Mansfield RE,et al.NMR spectroscopy as a tool for the rapid assessment of the conformation of GST-fusion proteins[J].Protein Sci,2008,17(9):1630-1635.
• [33]Bell MR,Engleka MJ,Malik A,et al.To fuse or not to fuse:what is your purpose[J].Protein Sci,2013[Epub ahead of print].