Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2.
Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2.
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细胞外环2中的保守残基调节Stachel介导的ADGRG2活化。
DOI：
10.1038/s41598-021-93577-y
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发表时间：
2021-07-07
影响因子：
4.6
通讯作者：
Balenga N
中科院分区：
文献类型：
--
作者：
Gad AA;Azimzadeh P;Balenga N
粘附性G蛋白偶联受体(AGPCR)家族的一些成员的激活机制是N-末端大片段的切割和解离以及随后N-末端短序列(Stachel)的揭开。然而，在同源Stachel激活aGPCRs过程中发挥作用的残基的身份在很大程度上仍不清楚。蛋白质序列比对显示，aGPCR家族所有33个成员的胞外环2(ECL2)中都有一段保守的残基。ADGRG2是一种孤儿aGPCR，在男性生育、尤文肉瘤细胞增殖和甲状旁腺细胞功能等方面发挥重要作用。我们以ADGRG2为模型，通过定点突变获得了ECL2中保守残基的突变体。我们发现ECL2中的色氨酸和异亮氨酸对于HEK293细胞的受体稳定性和表面表达是必不可少的。通过调节受体表面的表达水平，我们发现这些ECL2残基的突变破坏了Stachel介导的ADGRG2多条信号通路的激活。本研究对ECL2在Stachel介导的信号转导和ADGRG2降解中的作用提供了新的认识，为合理设计针对aGPCRs的治疗药物奠定了基础。
Cleavage and dissociation of a large N-terminal fragment and the consequent unmasking of a short sequence (Stachel) remaining on the N-terminus have been proposed as mechanisms of activation of some members of the adhesion G protein-coupled receptor (aGPCR) family. However, the identity of residues that play a role in the activation of aGPCRs by the cognate Stachel remains largely unknown. Protein sequence alignments revealed a conserved stretch of residues in the extracellular loop 2 (ECL2) of all 33 members of the aGPCR family. ADGRG2, an orphan aGPCR, plays a major role in male fertility, Ewing sarcoma cell proliferation, and parathyroid cell function. We used ADGRG2 as a model aGPCR and generated mutants of the conserved residues in the ECL2 via site-directed mutagenesis. We show that tryptophan and isoleucine in the ECL2 are essential for receptor stability and surface expression in the HEK293 cells. By adjusting the receptor surface expression levels, we show that mutation of these residues of ECL2 ablates the Stachel-mediated activation of multiple signaling pathways of ADGRG2. This study provides a novel understanding of the role of the ECL2 in Stachel-mediated signaling and degradation of ADGRG2, which may lay the foundation for the rational design of therapeutics to target aGPCRs.
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影响因子：
16.2
作者：
Petersen, Sarah C.;Luo, Rong;Liebscher, Ines;Giera, Stefanie;Jeong, Sung-Jin;Mogha, Amit;Ghidinelli, Monica;Feltri, M. Laura;Schoeneberg, Torsten;Piao, Xianhua;Monk, Kelly R.
通讯作者：
Monk, Kelly R.
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13.8
作者：
Ellaithy A;Gonzalez-Maeso J;Logothetis DA;Levitz J
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Levitz J
影响因子：
7.3
作者：
Paavola KJ;Sidik H;Zuchero JB;Eckart M;Talbot WS
通讯作者：
Talbot WS
影响因子：
16.8
作者：
Ehrenmann, Janosch;Schoeppe, Jendrik;Plueckthun, Andreas
通讯作者：
Plueckthun, Andreas
影响因子：
64.5
作者：
Inoue, Asuka;Raimondi, Francesco;Russell, Robert B.
通讯作者：
Russell, Robert B.