FAM237A, rather than peptide PEN and proCCK56-63, binds to and activates the orphan receptor GPR83
FAM237A, rather than peptide PEN and proCCK56-63, binds to and activates the orphan receptor GPR83
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DOI：
10.1111/febs.16765
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发表时间：
2023-03-07
期刊：
影响因子：
5.4
通讯作者：
Guo, Zhan-Yun
中科院分区：
文献类型：
--
作者：
Li, Hao-Zheng;Wang, Ya-Fen;Guo, Zhan-Yun
G蛋白偶联受体83（GPR 83）主要在大脑中表达，并参与能量代谢和一些焦虑相关行为的调节。最近，PCSK 1 N/proSAAS衍生肽PEN、前胆囊收缩素衍生肽proCCK 56 -63和具有序列相似性的家族237成员A（FAM 237 A）都被报道为GPR 83的有效激动剂。然而，这些结果尚未被其他实验室重现，因此GPR 83仍然是官方的孤儿受体。肽PEN和proCCK 56 -63具有序列相似性;然而，它们与FAM 237 A完全不同。为了鉴定其实际配体，在本研究中，我们开发了基于NanoLuc二元技术（NanoBiT）的配体结合测定、基于荧光配体的可视化和基于NanoBiT的人GPR 83的β-抑制蛋白募集测定。使用这些测定，我们证明了成熟的人FAM 237 A可以以纳摩尔范围的亲和力结合GPR 83，并且可以在转染的人胚肾293 T细胞中以纳摩尔范围的效率激活该受体并诱导其内化。然而，使用这些测定，我们没有检测到PEN和proCCK 56 -63与GPR 83的任何相互作用。因此，我们的结果证实FAM 237 A是GPR 83的有效激动剂，但不支持PEN和proCCK 56 -63作为该受体的配体。阐明它们的配对为将来进一步研究脑特异性受体GPR 83和迄今为止很少研究的神经肽FAM 237 A的功能铺平了道路。
G protein-coupled receptor 83 (GPR83) is primarily expressed in the brain and is implicated in the regulation of energy metabolism and some anxiety-related behaviours. Recently, the PCSK1N/proSAAS-derived peptide PEN, the procholecystokinin-derived peptide proCCK56-63, and family with sequence similarity 237 member A (FAM237A) were all reported as efficient agonists of GPR83. However, these results have not yet been reproduced by other laboratories and thus GPR83 is still officially an orphan receptor. The peptide PEN and proCCK56-63 share sequence similarity; however, they are completely different from FAM237A. To identify its actual ligand(s), in the present study we developed NanoLuc Binary Technology (NanoBiT)-based ligand-binding assay, fluorescent ligand-based visualization, and NanoBiT-based beta-arrestin recruitment assay for human GPR83. Using these assays, we demonstrated that mature human FAM237A could bind to GPR83 with nanomolar range affinity, and could activate this receptor and induce its internalization with nanomolar range efficiency in transfected human embryonic kidney 293T cells. However, we did not detect any interaction of PEN and proCCK56-63 with GPR83 using these assays. Thus, our results confirmed that FAM237A is an efficient agonist of GPR83, but did not support PEN and proCCK56-63 as ligands of this receptor. Clarification of their pairing paves the way for further functional studies of the brain-specific receptor GPR83 and the so far rarely studied neuropeptide FAM237A in the future.