8VHF

Cryo-EM structure of GPR119-Gs-Nb35 complex with small molecule agonist MBX-2982

Summary for 8VHF

• Entry DOI: 10.2210/pdb8vhf/pdb
• EMDB information: 43236
• Descriptor: Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1, HiBiT, Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2, Nanobody35, ... (6 entities in total)
• Functional Keywords: cryoem, gpcr, orphan, g protein complex, active, agonist, membrane protein, diabetes, native mass spectrometry, membrane protein-signaling protein complex, membrane protein/signaling protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 5
• Total formula weight: 176952.00

Authors

Kim, D.-G.,Cherezov, V. (deposition date: 2024-01-01, release date: 2024-10-30, Last modification date: 2025-03-05)

Primary citation

Kim, D.,Liu, W.,Viner, R.,Cherezov, V.
Native mass spectrometry prescreening of G protein-coupled receptor complexes for cryo-EM structure determination.
Structure, 32:2206-2219.e4, 2024

PubMed Abstract: G protein-coupled receptors (GPCRs) are essential transmembrane proteins playing key roles in human health and disease. Understanding their atomic-level molecular structure and conformational states is imperative for advancing drug development. Recent breakthroughs in single-particle cryogenic electron microscopy (cryo-EM) have propelled the structural biology of GPCRs into a new era. Nevertheless, the preparation of suitable GPCR samples and their complexes for cryo-EM analysis remains challenging due to their poor stability and highly dynamic nature. Here, we present our online buffer exchange-native MS method combined with Direct Mass Technology (OBE-nMS+DMT) which facilitates high-throughput analysis and guides sample preparation. We applied this method to optimize the GPR119-G complex sample prior to cryo-EM analysis, leading to a 3.51 Å resolution structure from only 396 movies collected on a 200 kV Glacios. This study suggests that the OBE-nMS+DMT method emerges as a powerful tool for prescreening sample conditions in cryo-EM studies of GPCRs and other membrane protein complexes.

PubMed: 39471802
DOI: 10.1016/j.str.2024.10.004

Experimental method

ELECTRON MICROSCOPY (3.51 Å)