Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins

	Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins
	Mukherjee, Somnath 1; Erramilli, Satchal K 1; Ammirati, Mark 2; Alvarez, Frances J D 2; Fennell, Kimberly F 2; Purdy, Michael D 3; Skrobek, Blazej M 1; Radziwon, Katarzyna 1; Coukos, John 1; Kang, Yanyong 6
刊名	NATURE COMMUNICATIONS
	2020
卷号	11 期号:1
ISSN号	2041-1723
英文摘要	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.
资助项目	[National Institutes of Health] ; [CHEETAH Collaborative Development Program] ; [Pfizer Inc.]
语种	英语
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/298607]
专题	中国科学院上海药物研究所
作者单位	1.University of Chicago 2.Pfizer 3.University of Virginia 4.University of Munich 5.University of Wisconsin System 6.Van Andel Institute 7.Takeda San Diego Inc 8.California Institute of Technology 9.Shanghai Institute of Materia Medica, Chinese Academy of Sciences
推荐引用方式 GB/T 7714	Mukherjee, Somnath,Erramilli, Satchal K,Ammirati, Mark,et al. Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins[J]. NATURE COMMUNICATIONS,2020,11(1).
APA	Mukherjee, Somnath.,Erramilli, Satchal K.,Ammirati, Mark.,Alvarez, Frances J D.,Fennell, Kimberly F.,...&Kossiakoff, Anthony A.(2020).Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins.NATURE COMMUNICATIONS,11(1).
MLA	Mukherjee, Somnath,et al."Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins".NATURE COMMUNICATIONS 11.1(2020).