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PLC-β3与syntrophin蛋白的相互作用 被引量:1

The interaction between PLC-β3 and syntrophin
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摘要 目的探索并鉴定PLC-β3与PDZ蛋白syntrophin的相互作用,为进一步研究PLC-β3在神经系统信号传导通路中的作用提供线索。方法制备GST-PLC-β3-CT融合蛋白,利用GST pull down的方法,检测PLC-β3-CT与β2-syntrophin和γ2-syntrophin的PDZ结构域的相互作用。结果构建重组质粒pGEX-PLC-β3-CT,在大肠杆菌BL21中成功表达融合蛋白后,利用GST pull down的方法证实了PLC-β3-CT可以与β2-syntrophin而不是γ2-syntrophin的PDZ结构域相互作用。结论 PLC-β3与β2-syntrophin相互作用,为研究完整的PLC-β3与β2-syntrophin的相互作用以及这种相互作用在神经系统信号网络中的功能提供了线索。 Objective To identify the interaction between PLC-β3 and the PDZ domain-containing protein syntrophin,and to find the underlying roles of PLC-β3 in signaling pathway of the nervous system.Methods The carboxyl-terminal of PLC-β3(PLC-β3-CT) was fused to GST protein by gene cloning.The resultant protein was used for examining interactions of PLC-β3-CT with the PDZ-domain of β2-syntrophin and γ2-syntrophin respectively by GST pull-down assay.Results PLC-β3-CT cDNA was cloned into the expression vector pGEX-4T-1 successfully,and the GST fusion protein was overexpressed and purified in an E.coli system.GST pull-down experiments revealed that the PDZ domain of β2-syntrophin,but not γ2-syntrophin was robustly pulled down by the carboxyl-terminal of PLC-β3.Conclusion These results provide a strong evidence for further studying the intracellular association of the PLC-β3 and β2-syntrophin and for exploring their potential physiological significances in signaling pathway of the nervous system.
出处 《基础医学与临床》 CSCD 北大核心 2010年第7期708-713,共6页 Basic and Clinical Medicine
基金 国家自然基金(30772573 30973405) 北京市自然基金(7082009) 北京市教育委员会科技发展计划重点项目(KZ201010025026) 北京市"创新人才建设计划"项目
关键词 PLC-β3 SYNTROPHIN 蛋白相互作用 PLC-β3 syntrophin protein interaction
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  • 1Pann GS, Jae IP, Lucia M, et al. Multiple roles of phosphoinositide-specific phospholipase C Isozymes [ J ]. Biochem Mol Bid Rep, 2008, 41(6):415 -434.
  • 2Resham B, Junhye K, Li Xiujuan, et al. Distinct role of PLC-β3 in VEGF-mediated directional migration and vascular sprouting[J]. J Cell Sci, 2009, 122(7): 1025 - 1034.
  • 3van Zeijl L, Ponsioen B, Ben Giepmans NG, et al. Regulation of connexin-43 gap junctional communication by phosphatidylinositol-4,5-bisphosphate [ J ]. J Cell Biolo, 2007, 177(5) :881 -891.
  • 4Alberto M. Phospholipase C beta 3 is a key component in the Gbetagamma/PKCeta/PKD-mediated regulation of trans- Golgi network to plasma membrane transport [ J ]. Biochem J, 2007, 406(1):157-165.
  • 5Jong IH, Hyeon SK, Jae RL, et al. The interaction of phospholipase C-133 with Shank2 regulates mGluR-mediated calcium signal [ J ]. J Biol Chem, 2005, 280 ( 13 ) : 12467 - 12473.
  • 6Yong SO, Nam W J, Jung WC, et al. NHERF2 specifically interacts with LPA2 receptor and defines the specificity and efficiency of receptor-mediated phospholipase C-β3 activation[J]. Mol Cell Biolo, 2004, 24(11) : 5069-5079.
  • 7Bohm SV, Constantinou P, Tan S, et al. Profound human/ mouse differences in alpha-dystrobrevin isoforms: a novel syntrophin-binding site and promoter missing in mouse and rat[J]. BMC Siol, 2009,7(85):1 -12.
  • 8Hogan A, Shepherd L, Chabot J, et al. Interaction of gamma 1-syntrophin with diacylglycerol kinase-zeta. Regulation of nuclear localization by PDZ interactions [ J ]. J Biol Chem, 2001, 276(28):26526-26533.
  • 9Ou Y, Strege P, Miller SM, et al. Syntrophin gamma 2 regulates SCN5A gating by a PDZ domain-mediated interaction[J]. J Biol Chem, 2003, 278(3):1915 -1923.
  • 10Leonoudakis D, Conti LR, Anderson S, et al. Protein tragicking and anchoring complexes revealed by proteomic analysis of inward rectifier potassium channel (Kir21x)-associated proteins [ J ]. J Biol Chem, 2004, 279 (21) : 22331 - 22346.

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  • 5Finn RD et al. Rainbow tags: a visual tag system for recombinant protein expression and purification. Biotechniques, 2005, 38:387-388.
  • 6Butt TR et al. Ubiquitin fusion augments the yield of cloned gene products E. eoli. Proc Natl Acad Sci USA, 1989, 86: 2540- 2544.
  • 7Korf U et al. Large-scale protein expression for proteome research. Proteomics, 2005, 5:3571-3580.
  • 8Chopra AK et al. Improved synthesis of Salmonedlla typhimunium enterotoxin using gene fusion expression system. Gene, 1994, 144: 81-85.
  • 9Chayen NE et al. Turning protein crystallization from an art into a science. Curr Opin Struct Biol, 2004, 14:577-583.
  • 10De Marco Vet al. The solubility and stability of recombinant proteins are increased by their fusion to NusA. Biochem Biophys Res Commun, 2004, 322:766-771.

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