Please use this identifier to cite or link to this item:
Title: An optimized method for 15N R1 relaxation rate measurements in non-deuterated proteins
Author: Gairí Tahull, Margarida
Dyachenko, Andrey
González, M. Teresa
Feliz Rodenas, Miguel
Pons Vallès, Miquel
Giralt Lledó, Ernest
Keywords: Proteïnes
Relaxació (Física nuclear)
Relaxation (Nuclear physics)
Issue Date: 4-Jun-2015
Publisher: Springer Verlag
Abstract: 15N longitudinal relaxation rates are extensively used for the characterization of protein dynamics; however, their accurate measurement is hindered by systematic errors. 15N CSA/1H-15N dipolar cross-correlated relaxation (CC) and amide proton exchange saturation transfer from water protons are the two main sources of systematic errors in the determination of 15N R1 rates through 1H-15N HSQC-based experiments. CC is usually suppressed through a train of 180° proton pulses applied during the variable 15N relaxation period (T), which can perturb water magnetization. Thus CC cancellation is required in such a way as to minimize water saturation effects. Here we examined the level of water saturation during the T period caused by various types of inversion proton pulses to suppress CC: (I) amide-selective IBURP-2; (II) cosine-modulated IBURP-2; (III) Watergate-like blocks; and (IV) non-selective hard. We additionally demonstrate the effect of uncontrolled saturation of aliphatic protons on 15N R1 rates. In this paper we present an optimized pulse sequence that takes into account the crucial effect of controlling also the saturation of the aliphatic protons during 15N R1 measurements in non-deuterated proteins. We show that using cosine-modulated IBURP-2 pulses spaced 40 ms to cancel CC in this optimized pulse program is the method of choice to minimize
Note: Versió postprint del document publicat a:
It is part of: Journal of Biomolecular NMR, 2015, vol. 62, num. 2, p. 209-220
Related resource:
ISSN: 0925-2738
Appears in Collections:Articles publicats en revistes (Química Inorgànica i Orgànica)

Files in This Item:
File Description SizeFormat 
653488.pdf1.61 MBAdobe PDFView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.