8XGR

ETB-eGt complex bound to endothelin-1

Summary for 8XGR

• Entry DOI: 10.2210/pdb8xgr/pdb
• EMDB information: 38330
• Descriptor: Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2,eGt-alpha, Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1, Camelid antibody VHH fragment, ... (5 entities in total)
• Functional Keywords: signaling protein, peptide binding protein-immune system complex, peptide binding protein/immune system
• Biological source: Bos taurus (cattle); More
• Total number of polymer chains: 5
• Total formula weight: 176854.69

Authors

Oshima, H.S.,Sano, F.K.,Akasaka, H.,Iwama, A.,Shihoya, W.,Nureki, O. (deposition date: 2023-12-15, release date: 2024-04-03, Last modification date: 2025-07-02)

Primary citation

Oshima, H.S.,Sano, F.K.,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., 693:149361-149361, 2024

PubMed Abstract: 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 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 complexes remain both challenging and resource-intensive. By employing eGα, which exhibits binding affinity to modified nanobody Nb35, the cryo-EM structure of Rhodopsin-eGα complex was previously reported. Using this modified G protein, we determined the structure of the ET-eG complex bound to the modified Nb35. The determined structure of ET receptor was the same as the previously reported ET-G complex, and the resulting dataset demonstrated significantly improved anisotropy. This modified G protein will be utilized for the structural determination of other GPCR-G complexes.

PubMed: 38128244
DOI: 10.1016/j.bbrc.2023.149361

Experimental method

ELECTRON MICROSCOPY (3.2 Å)