Journal of Biomolecular NMR

, Volume 73, Issue 10–11, pp 641–650 | Cite as

Revisiting 1HN CPMG relaxation dispersion experiments: a simple modification can eliminate large artifacts

  • Tairan Yuwen
  • Lewis E. KayEmail author


Carr–Purcell–Meiboom–Gill relaxation dispersion experiments are commonly used to probe biomolecular dynamics on the millisecond timescale. The simplest experiment involves using backbone 15N spins as probes of motion and pulse sequences are now available for providing accurate dispersion profiles in this case. In contrast, 1H-based experiments recorded on fully protonated samples are less common because of difficulties associated with homonuclear scalar couplings that can result in transfer of magnetization between coupled spins, leading to significant artifacts. Herein we examine a version of the 1HN CPMG experiment that has been used in our laboratory where a pair of CPMG pulse trains comprising non-selective, high power 1H refocusing pulses sandwich an amide selective pulse that serves to refocus scalar-coupled evolution by the end of the train. The origin of the artifacts in our original scheme is explained and a new, significantly improved sequence is presented. The utility of the new experiment is demonstrated by obtaining flat 1HN dispersion profiles in a protonated protein system that is not expected to undergo millisecond timescale dynamics, and subsequently by measuring profiles on a cavity mutant of T4 lysozyme that exchanges between a pair of distinct states, establishing that high quality data can be generated even for fully protonated samples.


Chemical exchange 1HN CPMG Amide protons Fully protonated proteins Invisible excited states Protein dynamics 



This work was supported by grants from the Canadian Institutes of Health Research (CIHR) and the Natural Sciences and Engineering Research Council of Canada (NSERC) to L.E.K. T.Y. acknowledges post-doctoral support from the CIHR. We thank Pramodh Vallurupalli for useful discussions about exchange dynamics in L99A T4L. L.E.K. holds a Canada Research Chair in Biochemistry.

Supplementary material

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Supplementary material 1 (PDF 663 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Departments of Molecular Genetics, Biochemistry and ChemistryUniversity of TorontoTorontoCanada
  2. 2.Program in Molecular MedicineHospital for Sick ChildrenTorontoCanada

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