Related ArticlesA CEST NMR experiment to obtain glycine 1H? chemical shifts in 'invisible' minor states of proteins.
J Biomol NMR. 2020 Jul 21;:
Authors: Tiwari VP, Vallurupalli P
Abstract
Chemical exchange saturation transfer (CEST) experiments are routinely used to study protein conformational exchange between a 'visible' major state and 'invisible' minor states because they can detect minor states with lifetimes varying from ~ 3 to ~ 100*ms populated to just ~ 0.5%. Consequently several 1H, 15N and 13C CEST experiments have been developed to study exchange and obtain minor state chemical shifts at almost all backbone and sidechain sites in proteins. Conspicuously missing from this extensive set of CEST experiments is a 1H CEST experiment to study exchange at glycine (Gly) 1H? sites as the existing 1H CEST experiments that have been designed to study dynamics in amide 1H-15N spin systems and methyl 13CH3 groups with three equivalent protons while suppressing 1H-1H NOE induced dips are not suitable for studying exchange in methylene 13CH2 groups with inequivalent protons. Here a Gly 1H? CEST experiment to obtain the minor state Gly 1H? chemical shifts is presented. The utility of this experiment is demonstrated on the L99A cavity mutant of T4 Lysozyme (T4L L99A) that undergoes conformational exchange between two compact conformers. The CEST derived minor state Gly 1H? chemical shifts of T4L L99A are in agreement with those obtained previously using CPMG techniques. The experimental strategy presented here can also be used to obtain methylene proton minor state chemical shifts from protein sidechain and nucleic acid backbone sites.
PMID: 32696193 [PubMed - as supplied by publisher]
A CEST NMR experiment to obtain glycine 1 H Ī± chemical shifts in ā??invisibleā?? minor states of proteins
A CEST NMR experiment to obtain glycine 1 H Ī± chemical shifts in ā??invisibleā?? minor states of proteins
Abstract
Chemical exchange saturation transfer (CEST) experiments are routinely used to study protein conformational exchange between a ā??visibleā?? major state and ā??invisibleā?? minor states because they can detect minor states with lifetimes varying fromā??~ā??3 toā??~ā??100Ā*ms populated to justā??~ā??0.5%. Consequently several 1H, 15N and 13C CEST experiments have been developed to study exchange and obtain minor state chemical shifts at...
A new class of CEST experiment based on selecting different magnetization components at the start and end of the CEST relaxation element: an application to 1 H CEST
A new class of CEST experiment based on selecting different magnetization components at the start and end of the CEST relaxation element: an application to 1 H CEST
Abstract
Chemical exchange saturation transfer (CEST) experiments are becoming increasingly popular for investigating biomolecular exchange dynamics with rates on the order of approximately 50ā??500Ā*sā??1 and a rich toolkit of different methods has emerged over the past few years. Typically, experiments are based on the evolution of longitudinal magnetization, or in some cases two-spin...
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01-19-2018 08:57 PM
[NMR paper] Transfer Rate Edited Experiment for the Selective Detection of Chemical Exchange via Saturaion Transfer (TRE-CEST)
Transfer Rate Edited Experiment for the Selective Detection of Chemical Exchange via Saturaion Transfer (TRE-CEST)
Publication date: Available online 7 May 2015
Source:Journal of Magnetic Resonance</br>
Author(s): Joshua I. Friedman , Ding Xia , Ravinder R. Regatte , Alexej Jerschow</br>
Chemical Exchange Saturation Transfer (CEST) magnetic resonance experiments have become valuable tools in magnetic resonance for the detection of low concentration solutes with far greater sensitivity than direct detection methods. Accurate measures of rates of chemical exchange...
13 C Ī± CEST experiment on uniformly 13 C-labeled proteins
13 C Ī± CEST experiment on uniformly 13 C-labeled proteins
Abstract
A new HSQC-based 13CĪ± CEST pulse scheme is proposed, which is suitable for uniformly 13C- or 13C, 15N-labeled samples in either water or heavy water. Except for Thr and Ser residues, the sensitivity of this scheme for uniformly labeled samples is similar to that of the previous scheme for selectively 13CĪ±-labeled samples with 100Ā*% isotope enrichment. The experiment is demonstrated on an acyl carrier protein domain. Our 13CĪ± CEST data reveal that the minor state of the acyl...
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12-03-2014 04:05 PM
[NMR paper] Effects of J couplings and unobservable minor states on kinetics parameters extracted from CEST data
Effects of J couplings and unobservable minor states on kinetics parameters extracted from CEST data
Publication date: Available online 31 October 2014
Source:Journal of Magnetic Resonance</br>
Author(s): Yang Zhou , Daiwen Yang</br>
Chemical Exchange Saturation Transfer (CEST) experiments have emerged as a powerful tool for characterizing dynamics and sparse populated conformers of protein in slow exchanging systems. We show that J couplings and invisible minor states can cause systematic errors in kinetics parameters and chemical shifts extracted from CEST...
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11-14-2014 08:33 AM
[NMR paper] A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins.
A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins.
A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins.
J Biomol NMR. 2013 Dec 18;
Authors: Tugarinov V, Venditti V, Marius Clore G
Abstract
A methyl-detected 'out-and-back' NMR experiment for obtaining simultaneous correlations of methyl resonances of valine and isoleucine/leucine residues with backbone carbonyl chemical shifts,...
A CEST NMR experiment to obtain glycine 1H? chemical shifts in 'invisible' minor states of proteins.
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