The N-terminal HSDCIF motif is required for cell surface trafficking and dimerization of family B G protein coupled receptor PAC1.
The N-terminal HSDCIF motif is required for cell surface trafficking and dimerization of family B G protein coupled receptor PAC1.
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N 端 HSDCIF 基序是 B G 家族蛋白偶联受体 PAC1 的细胞表面运输和二聚化所必需的
DOI：
10.1371/journal.pone.0051811
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhang H
中科院分区：
文献类型：
--
作者：
Yu R;Guo X;Zhong J;Li M;Zeng Z;Zhang H
PAC 1是垂体腺苷酸环化酶激活多肽（PACAP）的首选受体，属于B类G蛋白偶联受体（GPCR），主要介导PACAP的作用。G蛋白偶联受体同/异聚化在受体折叠、成熟、运输和细胞表面表达中的重要作用已变得越来越明显。本研究采用双分子荧光互补（BiFC）和生物发光共振能量转移（BRET）技术首次证实了PAC 1的二聚化。使用缺失了HSDCIF基序的受体突变体来分析结构-活性关系，该结构集中于位于信号序列之后并且与PACAP（1-6）具有高度同源性的N-末端HSDCIF基序。荧光共聚焦显微镜观察显示，HSDCIF基序的缺失损害了PAC 1的细胞递送。BiFC、BRET和westernblot结果表明，缺失HSDCIF基序并将Cys残基替换为Ala，可导致受体二聚化的破坏。外源化学合成的寡肽HSDCIF（100 nmol/L）不仅能下调PAC 1的二聚化，诱导PAC 1的内化，而且能抑制稳定表达PAC 1的CHO细胞的增殖，降低PACAP对细胞活力的影响。这些结果表明，N端HSDCIF基序在PAC 1的转运和二聚化过程中起着关键作用，外源性寡肽HSDCIF对细胞信号转导、转运和PAC 1的二聚化有影响。
PAC1 is PACAP (pituitary adenylate cyclase-activating polypeptide) preferring receptor belonging to class B G protein coupled receptor (GPCR) mediating the most effects of PACAP. The important role of G protein coupled receptor homo/heteromerization in receptor folding, maturation, trafficking, and cell surface expression has become increasingly evident. The bimolecular fluorescence complementation (BiFC) and bioluminescence resonance energy transfer (BRET) assay were used in this research to confirm the dimerization of PAC1 for the first time. The structure-activity relationship focused on the N-terminal HSDCIF motif, which locates behind the signal sequence and has high homology with PACAP (1–6), was assayed using a receptor mutant with the deletion of the HSDCIF motif. The fluorescence confocal microscope observation showed that the deletion of the HSDCIF motif impaired the cell delivery of PAC1. The results of BiFC, BRET and westernblot indicated that the deletion of HSDCIF motif and the replacement of the Cys residue with Ala in HSDCIF motif resulted in the disruption of receptor dimerization. And the exogenous chemically synthesized oligopeptide HSDCIF (100 nmol/L) not only down-regulated the dimerization of PAC1, induced the internalization of PAC1, but also inhibited the proliferation of CHO cells expressing PAC1 stably and decreased the activity of PACAP on the cell viability. All these data suggested that the N-terminal HSDCIF motif played key role in the trafficking and the dimerization of PAC1, and the exogenous oligopeptide HSDCIF had effects on the cell signaling, trafficking and the dimerization of PAC1.
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影响因子：
3.6
作者：
Asmann, YW;Dong, MQ;Miller, LJ
通讯作者：
Miller, LJ
DOI：
10.1111/j.1749-6632.2008.03630.x
发表时间：
2009-01-01
期刊：
TRENDS IN COMPARATIVE ENDOCRINOLOGY AND NEUROBIOLOGY
影响因子：
--
作者：
Boronkai, A.;Brubel, R.;Reglodi, D.
通讯作者：
Reglodi, D.
影响因子：
3.5
作者：
OKAZAKI, K;KIMURA, C;FUJINO, M
通讯作者：
FUJINO, M
DOI：
10.1111/j.1432-1033.1992.tb17043.x
发表时间：
1992-07-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
ROBBERECHT, P;GOURLET, P;CHRISTOPHE, J
通讯作者：
CHRISTOPHE, J
影响因子：
5.2
作者：
Roed SN;Orgaard A;Jorgensen R;De Meyts P
通讯作者：
De Meyts P