6CRK

Heterotrimeric G-protein in complex with an antibody fragment

Summary for 6CRK

• Entry DOI: 10.2210/pdb6crk/pdb
• Descriptor: Guanine nucleotide-binding protein G(i) subunit alpha-1, Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1, Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2, ... (7 entities in total)
• Functional Keywords: heterotrimeric g-protein antibody fragment, signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 4
• Total formula weight: 114428.43

Authors

Maeda, S.,Dawson, R.,Kobilka, B. (deposition date: 2018-03-19, release date: 2018-10-24, Last modification date: 2024-11-20)

Primary citation

Maeda, S.,Koehl, A.,Matile, H.,Hu, H.,Hilger, D.,Schertler, G.F.X.,Manglik, A.,Skiniotis, G.,Dawson, R.J.P.,Kobilka, B.K.
Development of an antibody fragment that stabilizes GPCR/G-protein complexes.
Nat Commun, 9:3712-3712, 2018

PubMed Abstract: Single-particle cryo-electron microscopy (cryo-EM) has recently enabled high-resolution structure determination of numerous biological macromolecular complexes. Despite this progress, the application of high-resolution cryo-EM to G protein coupled receptors (GPCRs) in complex with heterotrimeric G proteins remains challenging, owning to both the relative small size and the limited stability of these assemblies. Here we describe the development of antibody fragments that bind and stabilize GPCR-G protein complexes for the application of high-resolution cryo-EM. One antibody in particular, mAb16, stabilizes GPCR/G-protein complexes by recognizing an interface between Gα and Gβγ subunits in the heterotrimer, and confers resistance to GTPγS-triggered dissociation. The unique recognition mode of this antibody makes it possible to transfer its binding and stabilizing effect to other G-protein subtypes through minimal protein engineering. This antibody fragment is thus a broadly applicable tool for structural studies of GPCR/G-protein complexes.

PubMed: 30213947
DOI: 10.1038/s41467-018-06002-w

Experimental method

X-RAY DIFFRACTION (2 Å)