共 101 条
Site-specific labeling of proteins with NMR-active unnatural amino acids
被引:62
作者:
Jones, David H.
[1
]
Cellitti, Susan E.
[1
]
Hao, Xueshi
[1
]
Zhang, Qiong
[1
]
Jahnz, Michael
[2
,3
]
Summerer, Daniel
[2
,3
]
Schultz, Peter G.
[1
,2
,3
]
Uno, Tetsuo
[1
]
Geierstanger, Bernhard H.
[1
]
机构:
[1] Genom Inst Novartis Res Fdn, San Diego, CA 92121 USA
[2] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
[3] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
关键词:
Site-specific labeling;
Unnatural amino acids;
Spin label;
Metal chelator;
In-cell NMR;
IN-CELL NMR;
NUCLEAR-MAGNETIC-RESONANCE;
MOLECULAR-WEIGHT PROTEINS;
PARAMAGNETIC RELAXATION ENHANCEMENT;
CROSS-CORRELATED RELAXATION;
GLOBAL FOLD DETERMINATION;
ESCHERICHIA-COLI;
GENETIC-CODE;
MULTIDIMENSIONAL NMR;
DIRECTED SPIN;
D O I:
10.1007/s10858-009-9365-4
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions. The technology requires an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is added to the media while a TAG amber or frame shift codon specifies the incorporation site in the protein to be studied. These unnatural amino acids can be isotopically labeled and provide unique opportunities for site-specific labeling of proteins for NMR studies. In this perspective, we discuss these opportunities including new photocaged unnatural amino acids, outline usage of metal chelating and spin-labeled unnatural amino acids and expand the approach to in-cell NMR experiments.
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页码:89 / 100
页数:12
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