Lanthanoid tagging via an unnatural amino acid for protein structure characterization

doi:10.1007/s10858-017-0106-9

CORC > 武汉物理与数学研究所 > 中国科学院武汉物理与数学研究所 > 磁共振应用研究部

	Lanthanoid tagging via an unnatural amino acid for protein structure characterization
	Gu, Xin-Hua 2; Tang, Chun 1,2; Dong, Xu 2; Jiang, Wen-Xue 1,2
刊名	JOURNAL OF BIOMOLECULAR NMR
	2017-04-01
卷号	67 期号:4 页码:273-282
关键词	Unnatural Amino Acid Azide-alkyne Cycloaddition Pseudo-contact Shift Protein Structure Transient Protein Complex
DOI	10.1007/s10858-017-0106-9
文献子类	Article
英文摘要	Lanthanoid pseudo-contact shift (PCS) provides long-range structural information between a paramagnetic tag and protein nuclei. However, for proteins with native cysteines, site-specific attachment may only utilize functional groups orthogonal to sulfhydryl chemistry. Here we report two lanthanoid probes, DTTA-C3-yne and DTTA-C4-yne, which can be conjugated to an unnatural amino acid pAzF in the target protein via azide-alkyne cycloaddition. Demonstrated with ubiquitin and cysteine-containing enzyme EIIB, we show that large PCSs of distinct profiles can be generated for each tag/lanthanoid combination. The DTTA-based lanthanoid tags are associated with large magnetic susceptibility tensors owing to the rigidity of the tags. In particular, introduction of the DTTA-C3 tag affords intermolecular PCSs and enables structural characterization of a transient protein complex between ubiquitin and a UBA domain. Together, we have expanded the repertoire of paramagnetic tags and the applicability of paramagnetic NMR.
WOS关键词	PARAMAGNETIC RELAXATION ENHANCEMENT ; PSEUDOCONTACT SHIFTS ; PHOSPHOTRANSFERASE SYSTEM ; ESCHERICHIA-COLI ; NMR-SPECTROSCOPY ; WEAK ALIGNMENT ; GENETIC-CODE ; UBA DOMAIN ; TAGS ; COMPLEXES
WOS研究方向	Biochemistry & Molecular Biology ; Spectroscopy
语种	英语
WOS记录号	WOS:000400891000002
资助机构	Chinese Ministry of Science and Technology(2013CB910200 ; Chinese Ministry of Science and Technology(2013CB910200 ; National Natural Science Foundation of China(31225007 ; National Natural Science Foundation of China(31225007 ; K.C. Wong Education Foundation ; K.C. Wong Education Foundation ; Howard Hughes Medical Institute ; Howard Hughes Medical Institute ; 2016YFA0501200) ; 2016YFA0501200) ; 31400644) ; 31400644) ; Chinese Ministry of Science and Technology(2013CB910200 ; Chinese Ministry of Science and Technology(2013CB910200 ; National Natural Science Foundation of China(31225007 ; National Natural Science Foundation of China(31225007 ; K.C. Wong Education Foundation ; K.C. Wong Education Foundation ; Howard Hughes Medical Institute ; Howard Hughes Medical Institute ; 2016YFA0501200) ; 2016YFA0501200) ; 31400644) ; 31400644)
内容类型	期刊论文
源URL	[http://ir.wipm.ac.cn/handle/112942/11313]
专题	武汉物理与数学研究所_磁共振应用研究部
作者单位	1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, CAS Key Lab Magnet Resonance Biol Syst, Natl Ctr Magnet Resonance Wuhan,Collaborat Innova, Wuhan Inst Phys & Math,State Key Lab Magnet Reson, Wuhan 430071, Hubei, Peoples R China
推荐引用方式 GB/T 7714	Gu, Xin-Hua,Tang, Chun,Dong, Xu,et al. Lanthanoid tagging via an unnatural amino acid for protein structure characterization[J]. JOURNAL OF BIOMOLECULAR NMR,2017,67(4):273-282.
APA	Gu, Xin-Hua,Tang, Chun,Dong, Xu,&Jiang, Wen-Xue.(2017).Lanthanoid tagging via an unnatural amino acid for protein structure characterization.JOURNAL OF BIOMOLECULAR NMR,67(4),273-282.
MLA	Gu, Xin-Hua,et al."Lanthanoid tagging via an unnatural amino acid for protein structure characterization".JOURNAL OF BIOMOLECULAR NMR 67.4(2017):273-282.