Amino-acid type identification in 15N-HSQC spectra by combinatorial selective 15N-labelling

Journal of Biomolecular NMR
Peter S C WuGottfried Otting

Abstract

The efficiency of cell-free protein synthesis combined with combinatorial selective 15N-labelling provides a method for the rapid assignment of 15N-HSQC cross-peaks to the 19 different non-proline amino-acid types from five 15N-HSQC spectra. This strategy was explored with two different constructs of the C-terminal domain V of the tau subunit of the Escherichia coli DNA polymerase III holoenzyme, tauC16 and tauC14. Since each of the five 15N-HSQC spectra contained only about one third of the cross-peaks present in uniformly labelled samples, spectral overlap was much reduced. All 15N-HSQC cross-peaks of the backbone amides could be assigned to the correct amino-acid type. Availability of the residue-type information greatly assisted the evaluation of the changes in chemical shifts observed for corresponding residues in tauC16 vs. those in tauC14, and the analysis of the structure and mobility of the C-terminal residues present in tauC16 but not in tauC14.

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Citations

Jan 24, 2007·Journal of Biomolecular NMR·Takayoshi MatsudaTakanori Kigawa
Aug 9, 2008·Journal of Biomolecular NMR·Gottfried Otting
Apr 29, 2009·Journal of Biomolecular NMR·Xinying JiaGottfried Otting
Aug 15, 2009·Journal of Biomolecular NMR·Solmaz SobhanifarVolker Dötsch
Nov 6, 2010·Journal of Biomolecular NMR·Jun YokoyamaTakanori Kigawa
May 26, 2010·Proceedings of the National Academy of Sciences of the United States of America·Innokentiy MaslennikovSenyon Choe
Jul 9, 2010·New Biotechnology·Claire NoirotAnja Böckmann
Jun 18, 2014·Journal of the American Chemical Society·Rasmus LinserGerhard Wagner
Feb 18, 2010·Progress in Nuclear Magnetic Resonance Spectroscopy·Hak Jun KimCharles R Sanders
Mar 13, 2012·Chembiochem : a European Journal of Chemical Biology·Rodolfo M RasiaMichael J Plevin
Feb 3, 2012·Angewandte Chemie·Karin V LoschaGottfried Otting
Aug 22, 2015·Journal of Biomolecular NMR·Takuma KasaiTakanori Kigawa
Jul 31, 2012·Biochemical and Biophysical Research Communications·Ayako FurukawaMasato Katahira
Dec 3, 2014·Journal of Magnetic Resonance·Frank LöhrVolker Dötsch
Jan 25, 2011·Analytical Biochemistry·Jun YokoyamaTakanori Kigawa
Jul 8, 2016·Biochemistry·Paul J FockeFrancis I Valiyaveetil
Jun 25, 2017·Archives of Biochemistry and Biophysics·Takanori Kigawa

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