- EMDB-20863: CryoEM structure of human alpha4beta2 nicotinic acetylcholine rec... -
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データベース: EMDB / ID: EMD-20863
タイトル
CryoEM structure of human alpha4beta2 nicotinic acetylcholine receptor with varenicline in complex with anti-BRIL synthetic antibody BAK5
マップデータ
cytochrome B562-fused human alpha4beta2 nicotinic acetylcholine receptor in complex with fab fragments and bound to varenicline (focused on another alpha subunit complexed to a fab)
試料
複合体: human alpha4beta2 nicotinic acetylcholine receptor in complex fab fragments and bound to varenicline
タンパク質・ペプチド: chimera of soluble cytochrome b562 (BRIL) and neuronal acetylcholine receptor subunit alpha-4
ジャーナル: Nat Commun / 年: 2020 タイトル: Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins. 著者: Somnath Mukherjee / Satchal K Erramilli / Mark Ammirati / Frances J D Alvarez / Kimberly F Fennell / Michael D Purdy / Blazej M Skrobek / Katarzyna Radziwon / John Coukos / Yanyong Kang / ...著者: Somnath Mukherjee / Satchal K Erramilli / Mark Ammirati / Frances J D Alvarez / Kimberly F Fennell / Michael D Purdy / Blazej M Skrobek / Katarzyna Radziwon / John Coukos / Yanyong Kang / Przemysław Dutka / Xiang Gao / Xiayang Qiu / Mark Yeager / H Eric Xu / Seungil Han / Anthony A Kossiakoff / 要旨: 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 ...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.
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注釈
cytochrome B562-fused human alpha4beta2 nicotinic acetylcholine receptor in complex with fab fragments and bound to varenicline (focused on another alpha subunit complexed to a fab)
cytochrome B562-fused human alpha4beta2 nicotinic acetylcholine receptor in complex with fab fragments and bound to varenicline (focused on an alpha subunit complexed to a fab)
cytochrome B562-fused human alpha4beta2 nicotinic acetylcholine receptor in complex with fab fragments and bound to varenicline (focused on an alpha subunit complexed to a fab)