Assignment strategy for fast relaxing signals: complete aminoacid identification in thulium substituted calbindin D 9K

Journal of Biomolecular NMR
Stéphane BalayssacMario Piccioli

Abstract

Paramagnetic proteins generally contain regions with diverse relaxation properties. Nuclei in regions far from the metal center may behave like those in diamagnetic proteins, but those closer to the metal experience rapid relaxation with accompanying line broadening. We have used a set of NMR experiments optimized to capture data from these various concentric regions in assigning the signals from a paramagnetic Calbindin D 9K derivative in which one of the two calcium ions has been replaced by thulium(III). Normal double- and triple-resonance experiments with 1H detection were used in collecting data from nuclei in the diamagnetic-like region; these approaches identified signals from fewer than 50% of the amino acid residues (those with d > 17.5 A from thulium(III)). Paramagnetism-optimized two-dimensional NMR experiments with 1H detection were used in collecting data from nuclei in the next nearer region (d > 15 A). Standard (d > 14 A) and optimized (d > 9 A) 13C direct-detection experiments were used to capture data from nuclei in the next layer. Finally nuclei closest to the metal were detected by one-dimensional 13C (d > 5 A) and one-dimensional 15N data collection (d > 4.2 A). NMR signals were assigned on the basis of thro...Continue Reading

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Citations

Apr 19, 2013·Proceedings of the National Academy of Sciences of the United States of America·Lucia BanciJulia Winkelmann
Jan 31, 2012·Journal of Molecular Biology·Christophe SchmitzThomas Huber
May 23, 2009·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Isabella C Felli, Bernhard Brutscher
Oct 3, 2007·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Michael John, Gottfried Otting
Jan 16, 2010·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Wolfgang BermelRoberta Pierattelli
Oct 3, 2009·Journal of the American Chemical Society·Wolfgang BermelRoberta Pierattelli
Oct 7, 2016·Physical Chemistry Chemical Physics : PCCP·Elizaveta A Suturina, Ilya Kuprov
Dec 24, 2017·Nature Communications·Bassam K KudhairJeffrey Green
Jul 17, 2008·Dalton Transactions : an International Journal of Inorganic Chemistry·Ivano BertiniRoberta Pierattelli
May 14, 2020·PLoS Computational Biology·Eric WrightSunita Chandrasekaran
Jul 28, 2020·Journal of Biomolecular NMR·Michele InverniciMario Piccioli
Mar 12, 2017·Progress in Nuclear Magnetic Resonance Spectroscopy·Christoph Nitsche, Gottfried Otting
Nov 23, 2006·Journal of the American Chemical Society·Stéphane BalayssacMario Piccioli
Oct 13, 2007·Journal of the American Chemical Society·Michael JohnGottfried Otting
Jan 18, 2007·Journal of the American Chemical Society·Michael JohnGottfried Otting

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