咪唑啉1型受体候选蛋白信号转导的研究
发布时间：2018-07-28 12:39
【摘要】：咪唑啉1型受体(imidazoline-1 receptor,I1R)是近年发现的一种新型受体,能够介导许多生理作用。2000年Piletz教授成功地克隆出了咪唑啉受体抗体选择性蛋白(imidazoline receptor antisera selected protein,IRAS),为进一步研究I1R的功能和信号转导过程奠定了基础。 本研究应用本实验室已经建立的稳定转染IRAS的细胞(CHO-IRAS),对IRAS偶联的信号转导途径进行了初步研究。经典I1R激动剂莫索尼定、利美尼定及其内源性配体胍丁胺激活IRAS后并不能显著提高CHO-IRAS细胞的[~(35)S]-GTPγS结合量,表明IRAS不与G蛋白偶联。利美尼定、莫索尼定及胍丁胺激活IRAS后时间及浓度依赖性地引起磷脂酰胆碱特异性磷脂酶C(phosphatidylcholine selectivephospholipase C,PC-PLC)活性升高,且这一作用能被PC-PLC特异性拮抗剂D609及I1R选择性拮抗剂依法克生所抑制。上述药物在引起PC-PLC活性升高的同时,也能显著促进甘油二酯(diacylglyceride,DAG)含量增加。利美尼定、莫索尼定及胍丁胺都能引起细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)磷酸化水平发生改变。利美尼定和莫索尼定可以浓度依赖性地升高ERK磷酸化水平;但胍丁胺对ERK磷酸化有双重作用,低浓度时(1 nM-10 nM)降低ERK磷酸化水平,高浓度时(10 μM-100μM)促进ERK磷酸化。利美尼定、莫索尼定及胍丁胺对ERK磷酸化的作用能被PC-PLC特异性拮抗剂D609和I1R拮抗剂依法克生所抑制。 本研究首次直接证实IRAS不是G蛋白偶联受体。IRAS的信号转导途径可能是通过活化PC-PLC继而产生DAG,其后引起了丝裂原激活蛋白激酶(mitogen-activated protein kinase,MAPK)酶促级联反应过程。本研究为进一步阐明IRAS作为I1R候选蛋白而产生相应功能的分子机制提供了重要线索。
[Abstract]:Imidazoline type 1 receptor (I1R) is a new type of receptor found in recent years. In 2000, Professor Piletz successfully cloned Imidazoline receptor antibody selective protein (imidazoline receptor antisera selected protein (IRAs), which laid a foundation for the further study of I1R function and signal transduction process. In this study, we studied the signal transduction pathway of IRAS coupling using stable transfected IRAS cells (CHO-IRAS) established in our laboratory. Moxonidine, riminidine and its endogenous ligand agmatine did not significantly increase the binding capacity of [35S] -GTP 纬 S in CHO-IRAS cells, suggesting that IRAS was not coupled with G protein. The activity of phosphatidylcholine-specific phospholipase (C (phosphatidylcholine selectivephospholipase) was increased in a dose-and time-dependent manner after the activation of IRAS by laminidine, moxonidine and agmatine. This effect was inhibited by PC-PLC specific antagonist D609 and I1R selective antagonist Fokeson. The above drugs can increase the activity of PC-PLC and increase the content of diacylglyceride. Laminidine, moxonidine and agmatine can all cause changes in extracellular signal regulated kinase (extracellular signal-regulated kinase) phosphorylation. The level of ERK phosphorylation was increased in a concentration-dependent manner by nimenidine and moxonidine, but agmatine had a dual effect on ERK phosphorylation. At low concentration (1 nM-10 nm), ERK phosphorylation level was decreased, and ERK phosphorylation was promoted at high concentration (10 渭 M-100 渭 M). The phosphorylation of ERK by liminidine, moxonidine and agmatine was inhibited by PC-PLC specific antagonists D609 and I1R antagonists Fokeson. It is the first direct evidence that IRAS is not a G protein-coupled receptor. IRAs signal transduction pathway may be by activating PC-PLC to produce PC-PLC, and then induce the process of mitogen-activated protein kinase-activated protein kinase (mitogen-activated protein kinase) enzymatic cascade reaction. This study provides important clues for further elucidating the molecular mechanism of IRAS as a candidate protein for I1R.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33
本文编号：2150168
[Abstract]:Imidazoline type 1 receptor (I1R) is a new type of receptor found in recent years. In 2000, Professor Piletz successfully cloned Imidazoline receptor antibody selective protein (imidazoline receptor antisera selected protein (IRAs), which laid a foundation for the further study of I1R function and signal transduction process. In this study, we studied the signal transduction pathway of IRAS coupling using stable transfected IRAS cells (CHO-IRAS) established in our laboratory. Moxonidine, riminidine and its endogenous ligand agmatine did not significantly increase the binding capacity of [35S] -GTP 纬 S in CHO-IRAS cells, suggesting that IRAS was not coupled with G protein. The activity of phosphatidylcholine-specific phospholipase (C (phosphatidylcholine selectivephospholipase) was increased in a dose-and time-dependent manner after the activation of IRAS by laminidine, moxonidine and agmatine. This effect was inhibited by PC-PLC specific antagonist D609 and I1R selective antagonist Fokeson. The above drugs can increase the activity of PC-PLC and increase the content of diacylglyceride. Laminidine, moxonidine and agmatine can all cause changes in extracellular signal regulated kinase (extracellular signal-regulated kinase) phosphorylation. The level of ERK phosphorylation was increased in a concentration-dependent manner by nimenidine and moxonidine, but agmatine had a dual effect on ERK phosphorylation. At low concentration (1 nM-10 nm), ERK phosphorylation level was decreased, and ERK phosphorylation was promoted at high concentration (10 渭 M-100 渭 M). The phosphorylation of ERK by liminidine, moxonidine and agmatine was inhibited by PC-PLC specific antagonists D609 and I1R antagonists Fokeson. It is the first direct evidence that IRAS is not a G protein-coupled receptor. IRAs signal transduction pathway may be by activating PC-PLC to produce PC-PLC, and then induce the process of mitogen-activated protein kinase-activated protein kinase (mitogen-activated protein kinase) enzymatic cascade reaction. This study provides important clues for further elucidating the molecular mechanism of IRAS as a candidate protein for I1R.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33
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1 李斐;咪唑啉1型受体候选蛋白信号转导的研究[D];中国人民解放军军事医学科学院;2005年
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