Native mass spectrometry prescreening of G protein-coupled receptor complexes for cryo-EM structure determination

  • Structure. 2024 Dec 5;32(12):2206-2219.e4. doi: 10.1016/j.str.2024.10.004.
Donggyun Kim  1 ,  Weijing Liu  2 ,  Rosa Viner  3 ,  Vadim Cherezov  4
Affiliations
  • 1. Bridge Institute, Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA; Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA.
  • 2. Thermo Fisher Scientific, 355 River Oaks Pkwy, San Jose, CA 95134, USA.
  • 3. Thermo Fisher Scientific, 355 River Oaks Pkwy, San Jose, CA 95134, USA. Electronic address: [email protected].
  • 4. Bridge Institute, Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA; Department of Chemistry, University of Southern California, Los Angeles, CA 90089, USA. Electronic address: [email protected].
PMID: 39471802 DOI: 10.1016/j.str.2024.10.004
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-Gs 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.

Keywords
Direct Mass Technology; G protein-coupled receptor; GPR119; cryo-EM; membrane protein; native mass spectrometry; online buffer exchange.
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