期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 296, 期 -, 页码 -出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1016/j.jbc.2021.100798
关键词
-
资金
- NYSTEM Stem Cell Biology training grant [C322560GG]
- DFG postdoctoral fellowship [STE 2843/1-1]
- T32 Cell Biology training Grant [T32GM136542]
- DFG [266022790 P4, 209933838 B6, 421152132]
- NIH/NINDS [R01 NS102665]
- NYSTEM (NY State Stem Cell Science) [IIRP C32595GG]
- NIH/NIBIB [R01 EB028774]
- DFG (German Research Foundation) FOR2149
- NYU Perlmutter Cancer Center Support Grant [P30CA016087]
- NYU Grossman School of Medicine
The GPR133 receptor is cleaved shortly after protein synthesis, leading to increased canonical signaling. The dissociation of NTF and CTF at the plasma membrane promotes GPR133 activation and downstream signaling.
GPR133 (ADGRD1), an adhesion G protein-coupled receptor (GPCR) whose canonical signaling activates GaS-mediated generation of cytosolic cAMP, has been shown to be necessary for the growth of glioblastoma (GBM), a brain malignancy. The extracellular N terminus of GPR133 is thought to be autoproteolytically cleaved into N-terminal and C-terminal fragments (NTF and CTF, respectively). However, the role of this cleavage in receptor activation remains unclear. Here, we used subcellular fractionation and immunoprecipitation approaches to show that the WT GPR133 receptor is cleaved shortly after protein synthesis and generates significantly more canonical signaling than an uncleavable point mutant GPR133 (H543R) in patient-derived GBM cultures and HEK293T cells. After cleavage, the resulting NTF and CTF remain noncovalently bound to each other until the receptor is trafficked to the plasma membrane, where we demonstrated NTF-CTF dissociation occurs. Using a fusion of the CTF of GPR133 and the N terminus of thrombin-activated human protease-activated receptor 1 as a controllable proxy system to test the effect of intramolecular cleavage and dissociation, we also showed that thrombin-induced cleavage and shedding of the human protease-activated receptor 1 NTF increased intracellular cAMP levels. These results support a model wherein dissociation of the NTF from the CTF at the plasma membrane promotes GPR133 activation and downstream signaling. These findings add depth to our understanding of the molecular life cycle and mechanism of action of GPR133 and provide critical insights that will inform therapeutic targeting of GPR133 in GBM.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据