Site-specific labeling of proteins with NMR-active unnatural amino acids

被引：0
作者：
David H. Jones
Susan E. Cellitti
Xueshi Hao
Qiong Zhang
Michael Jahnz
Daniel Summerer
Peter G. Schultz
Tetsuo Uno
Bernhard H. Geierstanger
机构：
[1] Genomics Institute of the Novartis Research Foundation,Department of Chemistry and the Skaggs Institute for Chemical Biology
[2] The Scripps Research Institute,undefined
来源：
Journal of Biomolecular NMR | 2010年 / 46卷
关键词：
Site-specific labeling; Unnatural amino acids; Spin label; Metal chelator; In-cell NMR;
D O I：
暂无
中图分类号：
学科分类号：
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：11
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  • [11] Assfalg M(2008)In vivo incorporation of unnatural amino acids to probe structure, dynamics, and ligand binding in a large protein by nuclear magnetic resonance spectroscopy J Am Chem Soc 130 9268-9281
  • [12] Banci L(2009)A facile system for encoding unnatural amino acids in mammalian cells Angew Chem Int Ed Engl 48 4052-4055
  • [13] Bertini I(1999)Impact of residual dipolar couplings on the accuracy of NMR structures determined from a minimal number of NOE restraints J Am Chem Soc 121 6513-6514
  • [14] Turano P(2007)Elucidating transient macromolecular interactions using paramagnetic relaxation enhancement Curr Opin Struct Biol 17 603-616
  • [15] Vasos PR(2002)A new spin on protein dynamics Trends Biochem Sci 27 288-295
  • [16] Banci L(2001)Evaluation of site-directed spin labeling for characterizing protein-ligand complexes using simulated restraints Biophys J 81 1275-1284
  • [17] Bertini I(1996)Use of F-19 NMR to probe protein structure and conformational changes Annu Rev Biophys Biomol Struct 25 163-195
  • [18] Cantini F(2002)FlgM gains structure in living cells Proc Natl Acad Sci USA 99 12681-12684
  • [19] D’Amelio N(2005)Site-specific in vivo labeling of proteins for NMR studies ChemBioChem 6 55-58
  • [20] Gaggelli E(2006)A genetically encoded photocaged tyrosine Angew Chem Int Ed Engl 45 2728-2731
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