6CBV

Crystal structure of BRIL bound to an affinity matured synthetic antibody.

Summary for 6CBV

• Entry DOI: 10.2210/pdb6cbv/pdb
• Descriptor: Heavy chain, Fab fragment, Light Chain, Fab fragment, BRIL, ... (7 entities in total)
• Functional Keywords: 4-helix bundle, fusion protein, synthetic antibody, fab fragment, fusion protein in complex with fab, immune system
• Biological source: Homo sapiens; More
• Total number of polymer chains: 3
• Total formula weight: 59759.49

Authors

Mukherjee, S.,Skrobek, B.,Kossiakoff, A.A. (deposition date: 2018-02-05, release date: 2019-02-06, Last modification date: 2024-11-06)

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

X-RAY DIFFRACTION (1.872 Å)