Optimal isotope labelling for NMR protein structure determinations

Nature
Masatsune KainoshoPeter Güntert

Abstract

Nuclear-magnetic-resonance spectroscopy can determine the three-dimensional structure of proteins in solution. However, its potential has been limited by the difficulty of interpreting NMR spectra in the presence of broadened and overlapping resonance lines and low signal-to-noise ratios. Here we present stereo-array isotope labelling (SAIL), a technique that can overcome many of these problems by applying a complete stereospecific and regiospecific pattern of stable isotopes that is optimal with regard to the quality and information content of the resulting NMR spectra. SAIL uses exclusively chemically and enzymatically synthesized amino acids for cell-free protein expression. We demonstrate for the 17-kDa protein calmodulin and the 41-kDa maltodextrin-binding protein that SAIL offers sharpened lines, spectral simplification without loss of information, and the ability to rapidly collect the structural restraints required to solve a high-quality solution structure for proteins twice as large as commonly solved by NMR. It thus makes a large class of proteins newly accessible to detailed solution structure determination.

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Citations

Sep 23, 2008·European Biophysics Journal : EBJ·Peter Güntert
Sep 12, 2006·Journal of Biomolecular NMR·Mikhail ReibarkhGerhard Wagner
Sep 12, 2007·Journal of Biomolecular NMR·Touraj Etezady-EsfarjaniKurt Wüthrich
Sep 3, 2008·Journal of Biomolecular NMR·Kit I TongToshiyuki Tanaka
Oct 2, 2008·Journal of Biomolecular NMR·Toshihiko SugikiHideo Takahashi
May 27, 2009·Journal of Biomolecular NMR·Chenyun GuoVitali Tugarinov
Aug 15, 2009·Journal of Biomolecular NMR·Solmaz SobhanifarVolker Dötsch
Aug 12, 2009·Journal of Biomolecular NMR·David H JonesBernhard H Geierstanger
Oct 1, 2009·Journal of Biomolecular NMR·Kun LuMichael F Summers
Sep 29, 2009·Journal of Biomolecular NMR·Amy M Ruschak, Lewis E Kay
Sep 15, 2010·Journal of Biomolecular NMR·Garima JaipuriaHanudatta S Atreya
Nov 6, 2010·Journal of Biomolecular NMR·Jun YokoyamaTakanori Kigawa
Dec 15, 2010·Journal of Biomolecular NMR·B KrishnarjunaHanudatta S Atreya
Mar 31, 2011·Journal of Biomolecular NMR·Teppei IkeyaPeter Güntert
Jul 13, 2011·Journal of Biomolecular NMR·Aleksandr B SahakyanMichele Vendruscolo
Feb 22, 2012·Journal of Biomolecular NMR·Daniel GottsteinPeter Güntert
Nov 30, 2012·Journal of Biomolecular NMR·Karin V Loscha, Gottfried Otting
Jan 19, 2013·Journal of Biomolecular NMR·Lena BuchnerPeter Güntert
Apr 16, 2013·Journal of Biomolecular NMR·Frederik HefkePeter Güntert
Oct 19, 2013·Journal of Biomolecular NMR·Julien OrtsPeter Güntert
Nov 23, 2013·Journal of Biomolecular NMR·Roman J LichteneckerRobert Konrat
Jun 25, 2010·Journal of Structural and Functional Genomics·Li ZhaoGaetano T Montelione
Apr 8, 2010·Quarterly Reviews of Biophysics·Masatsune Kainosho, Peter Güntert
Mar 18, 2011·Quarterly Reviews of Biophysics·Paul Guerry, Torsten Herrmann
Feb 2, 2013·Biochemistry·Paul J BarrettCharles R Sanders
May 14, 2010·Journal of the American Chemical Society·Tobias MadlMichael Sattler

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