6USF

CryoEM structure of human alpha4beta2 nicotinic acetylcholine receptor with varenicline in complex with anti-BRIL synthetic antibody BAK5

Summary for 6USF

• Entry DOI: 10.2210/pdb6usf/pdb
• Related: 6CBV
• EMDB information: 20863
• Descriptor: chimera of soluble cytochrome b562 (BRIL) and neuronal acetylcholine receptor subunit alpha-4, Neuronal acetylcholine receptor subunit beta-2, Fab heavy chain, ... (9 entities in total)
• Functional Keywords: acetylcholine receptor, nicotine, varenicline, anti-bril synthetic antibody, membrane protein, membrane protein-immune system complex, membrane protein/immune system
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 9
• Total formula weight: 347972.25

Authors

Alvarez, F.J.D.,Mukherjee, S.,Han, S.,Ammirati, M.,Kossiakoff, A.A. (deposition date: 2019-10-26, release date: 2020-04-29, Last modification date: 2024-11-13)

Primary citation

Mukherjee, S.,Erramilli, S.K.,Ammirati, M.,Alvarez, F.J.D.,Fennell, K.F.,Purdy, M.D.,Skrobek, B.M.,Radziwon, K.,Coukos, J.,Kang, Y.,Dutka, P.,Gao, X.,Qiu, X.,Yeager, M.,Eric Xu, H.,Han, S.,Kossiakoff, A.A.
Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins.
Nat Commun, 11:1598-1598, 2020

PubMed Abstract: We propose the concept of universal fiducials based on a set of pre-made semi-synthetic antibodies (sABs) generated by customized phage display selections against the fusion protein BRIL, an engineered variant of apocytochrome b562a. These sABs can bind to BRIL fused either into the loops or termini of different GPCRs, ion channels, receptors and transporters without disrupting their structure. A crystal structure of BRIL in complex with an affinity-matured sAB (BAG2) that bound to all systems tested delineates the footprint of interaction. Negative stain and cryoEM data of several examples of BRIL-membrane protein chimera highlight the effectiveness of the sABs as universal fiducial marks. Taken together with a cryoEM structure of sAB bound human nicotinic acetylcholine receptor, this work demonstrates that these anti-BRIL sABs can greatly enhance the particle properties leading to improved cryoEM outcomes, especially for challenging membrane proteins.

PubMed: 32221310
DOI: 10.1038/s41467-020-15363-0

Experimental method

ELECTRON MICROSCOPY (3.87 Å)