National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM 133504
米国
Other private
McKnight Foundation
Other private
Irma T. Hirschl/Monique Weill-Caulier Trust
引用
ジャーナル: Sci Adv / 年: 2025 タイトル: Structural determinants of G protein subtype selectivity at the serotonin receptor 5-HT1A. 著者: Audrey L Warren / Gregory Zilberg / Anwar Abbassi / Alejandro Abraham / Shifan Yang / Daniel Wacker / 要旨: Activation of the serotonin receptor 5-HT1A has been shown to regulate mood and cognition, making 5-HT1A an important target in the treatment of anxiety, depression, and psychosis. Although the ...Activation of the serotonin receptor 5-HT1A has been shown to regulate mood and cognition, making 5-HT1A an important target in the treatment of anxiety, depression, and psychosis. Although the receptor signals through inhibitory G proteins, more work is necessary to understand differences in transducer coupling and its relation to functional activity. To develop a molecular understanding of the differences underlying transducer coupling and activation, we performed structure-activity relationship studies of 5-HT1A with distinct G proteins. Through a combination of in vitro assays, we identified a potent partial agonist that selectively engages a G protein subtype. We further investigated the differences in G protein engagement at 5-HT1A with cryo-electron microscopy, determining structures of 5-HT1A bound to distinct ligands and G protein subtypes. Combined with subsequent structure-guided mutagenesis and signaling assays, our studies uncover both orthosteric and allosteric determinants of agonist-specific stimulation of distinct transducers.
名称: Soluble cytochrome b562,5-hydroxytryptamine receptor 1A タイプ: protein_or_peptide / ID: 3 詳細: N-terminally truncated 5-HT1A receptor with a flag and his tag followed by a TEV cleavage site and BRIL at the N-terminus. コピー数: 2 / 光学異性体: LEVO
分子 #4: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2
分子
名称: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2 タイプ: protein_or_peptide / ID: 4 詳細: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2 with a GSAGSAGSA linker connecting it to the N-terminus of Guanine nucleotide-binding protein G(z) subunit alpha コピー数: 1 / 光学異性体: LEVO
凍結剤: ETHANE / 装置: LEICA EM GP 詳細: Blot force 3 for 3-5 seconds was used and subsequent grids were screened for ice thickness prior to data collection..
詳細
Sample was mono disperse following gel filtration. The sample was immediately concentrated for CryoEM grid preparation.