The palmitoylation of the N-terminal extracellular Cys37 mediates the nuclear translocation of VPAC1 contributing to its anti-apoptotic activity.
The palmitoylation of the N-terminal extracellular Cys37 mediates the nuclear translocation of VPAC1 contributing to its anti-apoptotic activity.
复制标题
N 端胞外 Cys37 的棕榈酰化介导 VPAC1 的核转位，有助于其抗凋亡活性
DOI：
10.18632/oncotarget.17449
复制
发表时间：
2017-06-27
期刊：
影响因子：
--
通讯作者：
Zhou T
中科院分区：
文献类型：
--
作者：
Yu R;Liu H;Peng X;Cui Y;Song S;Wang L;Zhang H;Hong A;Zhou T
VPAC 1是B类G蛋白偶联受体（GPCR），由垂体腺苷酸环化酶激活多肽（PACAP）和血管活性肠肽（VIP）共享。成熟VPAC 1的N-末端胞外结构域中的第一个半胱氨酸（Cys 37）是不参与保守的分子内二硫键形成的游离Cys。为了研究该Cys 37在VPAC 1中的生物学作用，在中国仓鼠卵巢（CHO）细胞中分别稳定表达了野生型VPAC 1和Cys 37/Ala突变体（VPAC 1-C37/A）与增强型黄色荧光蛋白（EYFP）的融合蛋白。VPAC 1-EYFP和VPAC 1-C37/A-EYFP均能正常转运至质膜，表达VPAC 1-EYFP的CHO细胞对喜树碱（CPT）诱导的细胞凋亡的抑制活性高于表达VPAC 1-C37/A-EYFP的细胞，而VPAC 1-C37/A-CHO细胞的增殖活性高于VPAC 1-CHO细胞。共聚焦显微镜分析、免疫印迹和荧光定量分析结果表明，在VIP（0.1 nM）的刺激下，VPAC 1-EYFP发生了明显的核转位，而VPAC 1-C37/A-EYFP未发生核转位。酰基-生物素交换试验和基于点击化学的棕榈酰化试验首次证实了生物信息学分析预测的Cys 37的棕榈酰化。棕榈酰化抑制剂2-溴棕榈酸酯可显著抑制VPAC 1-EYFP的核转位，同时抑制其抗凋亡活性。这些结果表明，棕榈酰化的Cys 37在N-末端的细胞外结构域的VPAC 1介导的核转位的VPAC 1有助于其抗凋亡活性。这些发现首次揭示了脂质介导的VPAC 1核转位产生了一种新的抗凋亡信号通路，这可能有助于促进以VPAC 1为靶点的新药开发策略。
VPAC1 is class B G protein-coupled receptors (GPCR) shared by pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP). The first cysteine (Cys37) in the N-terminal extracellular domain of mature VPAC1 is a free Cys not involved in the formation of conserved intramolecular disulfide bonds. In order to investigate the biological role of this Cys37 in VPAC1, the wild-type VPAC1 and Cys37/Ala mutant (VPAC1-C37/A) were expressed stably as fusion proteins with enhanced yellow fluorescent protein (EYFP) respectively in Chinese hamster ovary (CHO) cells. Both VPAC1-EYFP and VPAC1-C37/A-EYFP trafficked to the plasma membrane normally, and CHO cells expressing VPAC1-EYFP displayed higher anti-apoptotic activity against camptothecin (CPT) induced apoptosis than the cells expressing VPAC1-C37/A-EYFP, while VPAC1-C37/A-CHO cells showed higher proliferative activity than VPAC1-CHO cells. Confocal microscopic analysis, western blotting and fluorescence quantification assay showed VPAC1-EYFP displayed significant nuclear translocation while VPAC1-C37/A-EYFP did not transfer into nucleus under the stimulation of VIP (0.1 nM). Acyl-biotin exchange assay and click chemistry-based palmitoylation assay confirmed for the first time the palmitoylation of Cys37, which has been predicted by bioinformatics analysis. And the palmitoylation inhibitor 2-bromopalmitate significantly inhibited the nuclear translocation of VPAC1-EYFP and its anti-apoptotic activity synchronously. These results indicated the palmitoylation of the Cys37 in the N-terminal extracellular domain of VPAC1 mediates the nuclear translocation of VPAC1 contributing to its anti-apoptotic activity. These findings reveal for the first time the lipidation-mediating nuclear translocation of VPAC1 produces a novel anti-apoptotic signal pathway, which may help to promote new drug development strategy targeting VPAC1.
登录
查看更多内容
DOI：
10.1083/jcb.200911143
发表时间：
2010-10-04
期刊：
The Journal of cell biology
影响因子：
--
作者：
Misaki R;Morimatsu M;Uemura T;Waguri S;Miyoshi E;Taniguchi N;Matsuda M;Taguchi T
通讯作者：
Taguchi T
影响因子：
2.3
作者：
Estrada, Rosendo;Wang, Lichun;Jala, Venkatakrishna R.;Lee, Jen-Fu;Lin, Cheng-Yon;Gray, Robert D.;Haribabu, Bodduluri;Lee, Menq-Jer
通讯作者：
Lee, Menq-Jer
影响因子：
--
作者：
Coleman DT;Gray AL;Kridel SJ;Cardelli JA
通讯作者：
Cardelli JA
影响因子：
3.1
作者：
Karacay, B;O'Dorisio, MS;Krahe, R
通讯作者：
Krahe, R
影响因子：
2
作者：
Fernandez-Martinez, Ana B.;Carmena, Maria J.;Sanchez-Chapado, Manuel
通讯作者：
Sanchez-Chapado, Manuel