PDGFRβ与β2-syntrophin蛋白相互作用的初步研究
首发时间:2007-12-12
摘要:目的 探索并鉴定PDGFRβ与PDZ蛋白β2-syntrophin新的结合作用以及该相互作用的分子基础,为进一步研究PDGFRβ相关信号转导通路的调节提供线索。方法 制备GST-PDGFRβ 羧基末端的融合蛋白,通过GST pull-down 技术,检测PDGFRβ-CT与β2-syntrophin的PDZ 结构域及完整的β2-syntrophin 蛋白的相互作用。结果 成功构建了重组质粒pGEX-PDGFRβ-CT。在大肠杆菌中成功表达融合蛋白后,通过体外蛋白质结合实验证实了PDGFRβ-CT 与β2-syntrophin的PDZ结构域和兔脑组织中内源性的β2-syntrophin蛋白可以相互作用。结论 PDGFRβ-CT 与β2-syntrophin 之间存在相互作用,该相互作用是由PDGFRβ 的羧基末端与β2-syntrophin 的PDZ 结构域结合而实现的, 此结果为研究PDGFRβ 介导的下游信号转导通路以及细胞特定调控受体功能的机制奠定了基础。
关键词: PDGFRβ β2-syntrophin 蛋白质相互作用
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Characterization of the novel interaction between PDGFRβ and β2-syntrophin
Abstract:Objective The aim of this study is to identify the novel interaction between the PDZ domain-containing protein β2-syntrophin and platelet-derived growth factor receptor-β (PDGFRβ), further to explore the underlying mechanism of PDGFRβ/β2-syntrophin association. Methods The carboxyl-terminal of PDGFRβ (PDGFRβ-CT) was fused to GST protein by gene cloning. The interactions of PDGFRβ-CT with PDZ-domain β2-syntrophin and full-lengh β2-syntrophin were characterized by GST pull down assay. Results PDGFRβ-CT was cloned into pGEX-2TK and GST- fusion protein was over-expressed and purified in E.coli system. GST pull down experiment revealed that carboxyl-terminal of PDGFRβ robustly pulled down PDZ domain of β2-syntrophin. The interaction with endogenous β2-syntrophin was confirmed in the native rabbit brain tissue. Conclusion PDGFRβ specifically interacts with β2-syntrophin through the carboxyl-terminal of PDGFRβ and the PDZ domain on β2-syntrophin,which may modulate PDGFRβ downstream signaling pathway .
Keywords: PDGFRβ β2-syntrophin protein interaction
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PDGFRβ与β2-syntrophin蛋白相互作用的初步研究
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