6CBV

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

6CBV の概要

• エントリーDOI: 10.2210/pdb6cbv/pdb
• 分子名称: Heavy chain, Fab fragment, Light Chain, Fab fragment, BRIL, ... (7 entities in total)
• 機能のキーワード: 4-helix bundle, fusion protein, synthetic antibody, fab fragment, fusion protein in complex with fab, immune system
• 由来する生物種: Homo sapiens; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 59759.49

構造登録者

Mukherjee, S.,Skrobek, B.,Kossiakoff, A.A. (登録日: 2018-02-05, 公開日: 2019-02-06, 最終更新日: 2023-10-04)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.872 Å)