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Facile measurement of 1H–15N residual dipolar couplings in larger perdeuterated proteins

Journal of Biomolecular NMR volume 48, pages 65–70 (2010)Cite this article

Abstract

We present a simple method, ARTSY, for extracting 1JNH couplings and 1H–15N RDCs from an interleaved set of two-dimensional 1H–15N TROSY-HSQC spectra, based on the principle of quantitative J correlation. The primary advantage of the ARTSY method over other methods is the ability to measure couplings without scaling peak positions or altering the narrow line widths characteristic of TROSY spectra. Accuracy of the method is demonstrated for the model system GB3. Application to the catalytic core domain of HIV integrase, a 36 kDa homodimer with unfavorable spectral characteristics, demonstrates its practical utility. Precision of the RDC measurement is limited by the signal-to-noise ratio, S/N, achievable in the 2D TROSY-HSQC spectrum, and is approximately given by 30/(S/N) Hz.

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Acknowledgments

We thank Alexander Maltsev for the sample of perdeuterated GB3 and for help in preparing the liquid crystalline solution for IN50–212 measurement, and Lishan Yao for the protonated GB3 mutant. This work was supported in part by the Intramural Research Program of the NIDDK, NIH, and by the Intramural AIDS-Targeted Antiviral Program of the Office of the Director, NIH.

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Affiliations

  1. Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, 20892-0520, USA

    Nicholas C. Fitzkee & Ad Bax

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  1. Nicholas C. Fitzkee
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  2. Ad Bax
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Correspondence to Ad Bax.

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Fitzkee, N.C., Bax, A. Facile measurement of 1H–15N residual dipolar couplings in larger perdeuterated proteins. J Biomol NMR 48, 65–70 (2010). https://doi.org/10.1007/s10858-010-9441-9

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Keywords

  • ARTSY
  • Catalytic core domain
  • HIV integrase
  • Quantitative J correlation
  • TROSY
  • RDC