Title

Quantitative comparison of errors in 15N transverse relaxation rates measured using various CPMG phasing schemes

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

2012-5

Document Type

Article

Subjects

Molecular Dynamics Simulation; Monte Carlo Method; Nitrogen Isotopes; Nuclear Magnetic Resonance, Biomolecular; Proteins

Disciplines

Biochemistry, Biophysics, and Structural Biology | Pharmacology, Toxicology and Environmental Health

Abstract

Nitrogen-15 Carr-Purcell-Meiboom-Gill (CPMG) transverse relaxation experiment are widely used to characterize protein backbone dynamics and chemical exchange parameters. Although an accurate value of the transverse relaxation rate, R(2), is needed for accurate characterization of dynamics, the uncertainty in the R(2) value depends on the experimental settings and the details of the data analysis itself. Here, we present an analysis of the impact of CPMG pulse phase alternation on the accuracy of the (15)N CPMG R(2). Our simulations show that R(2) can be obtained accurately for a relatively wide spectral width, either using the conventional phase cycle or using phase alternation when the r.f. pulse power is accurately calibrated. However, when the r.f. pulse is miscalibrated, the conventional CPMG experiment exhibits more significant uncertainties in R(2) caused by the off-resonance effect than does the phase alternation experiment. Our experiments show that this effect becomes manifest under the circumstance that the systematic error exceeds that arising from experimental noise. Furthermore, our results provide the means to estimate practical parameter settings that yield accurate values of (15)N transverse relaxation rates in the both CPMG experiments.

DOI of Published Version

10.1007/s10858-012-9621-x

Source

J Biomol NMR. 2012 May;53(1):13-23. Epub 2012 Apr 1. Link to article on publisher's site

Journal/Book/Conference Title

Journal of biomolecular NMR

Related Resources

Link to Article in PubMed

PubMed ID

22466935

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