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.
引用
收藏
页码:89 / 100
页数:12
相关论文
共 101 条
[1]   Lanthanide-induced pseudocontact shifts for solution structure refinements of macromolecules in shells up to 40 Å from the metal ion [J].
Allegrozzi, M ;
Bertini, I ;
Janik, MBL ;
Lee, YM ;
Lin, GH ;
Luchinat, C .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (17) :4154-4161
[2]   The unusually stable quaternary structure of human Cu,Zn-superoxide dismutase 1 is controlled by both metal occupancy and disulfide status [J].
Arnesano, F ;
Banci, L ;
Bertini, I ;
Martinelli, M ;
Furukawa, Y ;
O'Halloran, TV .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (46) :47998-48003
[3]   Superoxide dismutase folding/unfolding pathway: Role of the metal ions in modulating structural and dynamical features [J].
Assfalg, M ;
Banci, L ;
Bertini, I ;
Turano, P ;
Vasos, PR .
JOURNAL OF MOLECULAR BIOLOGY, 2003, 330 (01) :145-158
[4]   Human SOD1 before harboring the catalytic metal - Solution structure of copper-depleted, disulfide-reduced form [J].
Banci, L ;
Bertini, I ;
Cantini, F ;
D'Amelio, N ;
Gaggelli, E .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (04) :2333-2337
[5]   Utilization of site-directed spin labeling and high-resolution heteronuclear nuclear magnetic resonance for global fold determination of large proteins with limited nuclear overhauser effect data [J].
Battiste, JL ;
Wagner, G .
BIOCHEMISTRY, 2000, 39 (18) :5355-5365
[6]   PROTON RELAXATION TIMES IN PARAMAGNETIC SOLUTIONS EFFECTS OF ELECTRON SPIN RELAXATION [J].
BLOEMBERGEN, N ;
MORGAN, LO .
JOURNAL OF CHEMICAL PHYSICS, 1961, 34 (03) :842-&
[7]   Isotope-filtered NMR methods for the study of biomolecular structure and interactions [J].
Breeze, AL .
PROGRESS IN NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY, 2000, 36 (04) :323-372
[8]   Polychromatic selective population inversion for TROSY experiments with large proteins [J].
Bromek, K ;
Lee, D ;
Hauhart, R ;
Krych-Goldberg, M ;
Atkinson, JP ;
Barlow, PN ;
Pervushin, K .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (01) :405-411
[9]   In-cell NMR for protein-protein interactions (STINT-NMR) [J].
Burz, David S. ;
Dutta, Kaushik ;
Cowburn, David ;
Shekhtman, Alexander .
NATURE PROTOCOLS, 2006, 1 (01) :146-152
[10]   Mapping structural interactions using in-cell NMR spectroscopy (STINT-NMR) [J].
Burz, DS ;
Dutta, K ;
Cowburn, D ;
Shekhtman, A .
NATURE METHODS, 2006, 3 (02) :91-93