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Random coil chemical shift for intrinsically disordered proteins: effects of temperature and pH

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Abstract

Secondary chemical shift analysis is the main NMR method for detection of transiently formed secondary structure in intrinsically disordered proteins. The quality of the secondary chemical shifts is dependent on an appropriate choice of random coil chemical shifts. We report random coil chemical shifts and sequence correction factors determined for a GGXGG peptide series following the approach of Schwarzinger et al. (J Am Chem Soc 123(13):2970–2978, 2001). The chemical shifts are determined at neutral pH in order to match the conditions of most studies of intrinsically disordered proteins. Temperature has a non-negligible effect on the 13C random coil chemical shifts, so temperature coefficients are reported for the random coil chemical shifts to allow extrapolation to other temperatures. The pH dependence of the histidine random coil chemical shifts is investigated in a titration series, which allows the accurate random coil chemical shifts to be obtained at any pH. By correcting the random coil chemical shifts for the effects of temperature and pH, systematic biases of the secondary chemical shifts are minimized, which will improve the reliability of detection of transient secondary structure in disordered proteins.

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Acknowledgments

This work was supported by the EliteForsk programme (M.K.), The John and Birthe Meyer Foundation, the Carlsberg Foundation grant number 2008-01-0368 and The Danish Natural Research Council grant numbers 272-08-0500 (F.M.P). We thank Kamil Tamiola and Frans Mulder (University of Groningen) for sharing their data for comparison of the random coil data sets and Gitte Wolfsberg Haxholm and Birthe B. Kragelund for valuable discussions and critical comments to the manuscript.

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  1. Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200, København N, Denmark

    Magnus Kjaergaard, Søren Brander & Flemming M. Poulsen

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  1. Magnus Kjaergaard
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  2. Søren Brander
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  3. Flemming M. Poulsen
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Correspondence to Flemming M. Poulsen.

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Kjaergaard, M., Brander, S. & Poulsen, F.M. Random coil chemical shift for intrinsically disordered proteins: effects of temperature and pH. J Biomol NMR 49, 139–149 (2011). https://doi.org/10.1007/s10858-011-9472-x

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