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果蝇来源的GPCR Methuselah G蛋白偶联信号转导通路研究
引用本文:张静,张儒,叶晨立,谢欣. 果蝇来源的GPCR Methuselah G蛋白偶联信号转导通路研究[J]. 细胞生物学杂志, 2011, 0(8)
作者姓名:张静  张儒  叶晨立  谢欣
作者单位:上海市信号转导与疾病研究重点实验室同济大学生命科学与技术学院;中国科学院上海药物研究所国家新药筛选中心;
基金项目:科技部科研项目(No.2008DFB30150); 上海市科委科研(No.08431910100,No.09DZ2260100,No.2010CB944901,No.2011CB965104)资助项目~~
摘    要:Methuselah(MTH)是果蝇来源的GPCR中的一员,它的突变可延长果蝇平均寿命并提高果蝇对外界胁迫因素的耐受性。但目前对MTH在细胞水平的信号转导研究鲜有报道。该研究用稳定表达MTH的HEK293细胞株,对与该受体偶联的G蛋白选择性做了研究。首先,用免疫荧光染色、Western blot及钙流实验验证了MTH在HEK293/Myc-MTH细胞表面能稳定表达,且具有正常生物学活性;MTH受体被其配体N-stunted活化后所引起细胞内钙的上升不能被PTX预处理抑制,提示活化的MTH可能通过与Gq/11而非Gi/o蛋白相偶联;进一步研究发现,MTH激活后不显著改变细胞中的cAMP水平,表明MTH不与Gs和Gi/o相偶联;MTH被激活后可引起ERK磷酸化。这些结果提示:MTH可能是Gq/11蛋白的偶联受体,为进一步研究MTH的下游信号转导和生物学功能奠定了基础。

关 键 词:G蛋白偶联受体  Methuselah受体  G蛋白  信号转导  

Characterization of G Protein-coupled Signal Transduction of the Drosophila GPCR Methuselah
Zhang Jing,Zhang Ru,Ye Chenli. Characterization of G Protein-coupled Signal Transduction of the Drosophila GPCR Methuselah[J]. Chinese Journal of Cell Biology, 2011, 0(8)
Authors:Zhang Jing  Zhang Ru  Ye Chenli
Affiliation:Zhang Jing~(1,2),Zhang Ru~1,Ye Chenli~(1,Xie Xin~(1,2*) (1 Shanghai Key Laboratory of Signaling and Disease Research,Laboratory of Receptor-based Bio-medicine,School of Life Sciences and Technology,Tongji University,Shanghai 200092,China,2 State Key Laboratory of Drug Research,the National Center for Drug Screening,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203,China)
Abstract:Drosophila Methuselah(MTH) is a member of the G protein-coupled receptor(GPCR) super family.Mutation of MTH receptor leads to extended life span and resistance to various forms of stress in fruit fly. However,the signal transduction of MTH at cellular level has rarely been studied.Using HEK293 cell line which stably expresses MTH,we investigated its G protein-coupling preference.Firstly,we confirmed the expression and biological activity of MTH in HEK293 cells by using immunofluorescent staining,Western blo...
Keywords:GPCR  Methuselah  G protein  signal transduction  
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