8CXO

Cryo-EM structure of the unliganded mSMO-PGS2 in a lipidic environment

Summary for 8CXO

• Entry DOI: 10.2210/pdb8cxo/pdb
• EMDB information: 27062 27063
• Descriptor: Smoothened homolog, GlgA glycogen synthase chimera, CHOLESTEROL (2 entities in total)
• Functional Keywords: g-protein coupled receptor, smoothened, unliganded state, lipid system, membrane protein
• Biological source: Mus musculus (house mouse); More
• Total number of polymer chains: 1
• Total formula weight: 81254.83

Authors

Zhang, K.,Wu, H.,Hoppe, N.,Manglik, A.,Cheng, Y. (deposition date: 2022-05-22, release date: 2022-08-03, Last modification date: 2024-10-09)

Primary citation

Zhang, K.,Wu, H.,Hoppe, N.,Manglik, A.,Cheng, Y.
Fusion protein strategies for cryo-EM study of G protein-coupled receptors.
Nat Commun, 13:4366-4366, 2022

PubMed Abstract: Single particle cryogenic-electron microscopy (cryo-EM) is used extensively to determine structures of activated G protein-coupled receptors (GPCRs) in complex with G proteins or arrestins. However, applying it to GPCRs without signaling proteins remains challenging because most receptors lack structural features in their soluble domains to facilitate image alignment. In GPCR crystallography, inserting a fusion protein between transmembrane helices 5 and 6 is a highly successful strategy for crystallization. Although a similar strategy has the potential to broadly facilitate cryo-EM structure determination of GPCRs alone without signaling protein, the critical determinants that make this approach successful are not yet clear. Here, we address this shortcoming by exploring different fusion protein designs, which lead to structures of antagonist bound A adenosine receptor at 3.4 Å resolution and unliganded Smoothened at 3.7 Å resolution. The fusion strategies explored here are likely applicable to cryo-EM interrogation of other GPCRs and small integral membrane proteins.

PubMed: 35902590
DOI: 10.1038/s41467-022-32125-2

Experimental method

ELECTRON MICROSCOPY (3.7 Å)