Structural highlights
Publication Abstract from PubMed
Cryo-EM single particle analysis has recently facilitated the high-resolution structural determination of numerous GPCR-G complexes. Diverse methodologies have been devised with this trend, and in the case of GPCR-G(i) complexes, scFv16, an antibody that recognizes the intricate interface of the complex, has been mainly implemented to stabilize the complex. However, owing to their flexibility and heterogeneity, structural determinations of GPCR-G(i) complexes remain both challenging and resource-intensive. By employing eGalpha(t), which exhibits binding affinity to modified nanobody Nb35, the cryo-EM structure of Rhodopsin-eGalpha(t) complex was previously reported. Using this modified G protein, we determined the structure of the ET(B)-eG(t) complex bound to the modified Nb35. The determined structure of ET(B) receptor was the same as the previously reported ET(B)-G(i) complex, and the resulting dataset demonstrated significantly improved anisotropy. This modified G protein will be utilized for the structural determination of other GPCR-G(i) complexes.
Optimizing cryo-EM structural analysis of G(i)-coupling receptors via engineered G(t) and Nb35 application.,Oshima HS, Sano FK, Akasaka H, Iwama A, Shihoya W, Nureki O Biochem Biophys Res Commun. 2024 Jan 22;693:149361. doi: , 10.1016/j.bbrc.2023.149361. Epub 2023 Dec 7. PMID:38128244[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Oshima HS, Sano FK, Akasaka H, Iwama A, Shihoya W, Nureki O. Optimizing cryo-EM structural analysis of G(i)-coupling receptors via engineered G(t) and Nb35 application. Biochem Biophys Res Commun. 2024 Jan 22;693:149361. PMID:38128244 doi:10.1016/j.bbrc.2023.149361
