Site-Specific Incorporation of Genetically Encoded Photo-Crosslinkers Locates the Heteromeric Interface of a GPCR Complex in Living Cells.
Site-Specific Incorporation of Genetically Encoded Photo-Crosslinkers Locates the Heteromeric Interface of a GPCR Complex in Living Cells.
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DOI：
10.1016/j.chembiol.2020.07.006
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发表时间：
2020-10-15
影响因子：
8.6
通讯作者：
Gonzalez-Maeso, Javier
中科院分区：
文献类型：
--
作者：
Shah, Urjita H.;Toneatti, Rudy;Gaitonde, Supriya A.;Shin, Jong M.;Gonzalez-Maeso, Javier
G蛋白偶联受体（gpcr）是细胞信号传导的重要介质。尽管能够以单体形式激活G蛋白，但许多研究表明a类gpcr可能与二聚体/低聚体相关。这些GPCR复合体中单个原体的相对位置仍然是一个激烈争论的话题。我们之前报道了A类5-HT2A受体（5-HT2AR）和C类代谢性谷氨酸2受体（mGluR2）能够形成GPCR异复合体。通过在mGluR2的跨膜（TM）部分的特定位置引入可光激活的非天然氨基酸对叠氮多- l -苯丙氨酸（azF），我们描绘了在5-HT2AR-mGluR2络合物的异质界面上物理相互作用的残基。我们发现5-HT2AR在mGluR2的TM4细胞内末端与azF结合交联，而在TM1和TM4的其他位置未观察到交联。总之，这些发现为5-HT2AR-mGluR2复合物的结构安排提供了重要的见解。Shah等人利用可光激活的非天然氨基酸的位点特异性掺入，发现mGluR2的TM4细胞内端与5-HT2AR发生物理相互作用。这些发现为5-HT2AR-mGluR2复合物的结构安排提供了重要的见解。这种多功能的方法可以应用于研究蛋白质-蛋白质复合物。
G protein-coupled receptors (GPCRs) are critical mediators of cell signaling. Although capable of activating G proteins in a monomeric form, numerous studies reveal a possible association of class A GPCRs into dimers/oligomers. The relative location of individual protomers within these GPCR complexes remains a topic of intense debate. We previously reported that class A serotonin 5-HT2A receptor (5-HT2AR) and class C metabotropic glutamate 2 receptor (mGluR2) are able to form a GPCR heterocomplex. By introducing the photoactivatable unnatural amino acid p-azido-L-phenylalanine (azF) at selected individual positions along the transmembrane (TM) segments of mGluR2, we delineate the residues that physically interact at the heteromeric interface of the 5-HT2AR-mGluR2 complex. We show that 5-HT2AR crosslinked with azF incorporated at the intracellular end of mGluR2’s TM4, while no crosslinking was observed at other positions along TM1 and TM4. Together, these findings provide important insights into the structural arrangement of the 5-HT2AR-mGluR2 complex. Using site-specific incorporation of photoactivatable unnatural amino acids, Shah et al. identified that the intracellular end of TM4 of mGluR2 physically interacts with 5-HT2AR. These findings provide important insights into the structural arrangement of the 5-HT2AR-mGluR2 complex. This versatile method can be applied to study protein-protein complexes.
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影响因子：
16.6
作者：
Rannversson H;Andersen J;Sørensen L;Bang-Andersen B;Park M;Huber T;Sakmar TP;Strømgaard K
通讯作者：
Strømgaard K
DOI：
10.1146/annurev.pharmtox.011008.145533
发表时间：
2010
影响因子：
12.5
作者：
Niswender CM;Conn PJ
通讯作者：
Conn PJ
影响因子：
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作者：
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通讯作者：
Sakmar, Thomas P.
影响因子：
3.6
作者：
Lopez-Gimenez, Juan F.;Canals, Meritxell;Milligan, Graeme
通讯作者：
Milligan, Graeme
影响因子：
2.9
作者：
Ray-Saha, Sarmistha;Huber, Thomas;Sakmar, Thomas P.
通讯作者：
Sakmar, Thomas P.