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Quantitative analysis of backbone motion in proteins using MAS solid-state NMR spectroscopy

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Abstract

We present a comprehensive analysis of protein dynamics for a micro-crystallin protein in the solid-state. Experimental data include 15N T 1 relaxation times measured at two different magnetic fields as well as 1H–15N dipole, 15N CSA cross correlated relaxation rates which are sensitive to the spectral density function J(0) and are thus a measure of T 2 in the solid-state. In addition, global order parameters are included from a 1H,15N dipolar recoupling experiment. The data are analyzed within the framework of the extended model-free Clore–Lipari–Szabo theory. We find slow motional correlation times in the range of 5 and 150 ns. Assuming a wobbling in a cone motion, the amplitude of motion of the respective amide moiety is on the order of 10° for the half-opening angle of the cone in most of the cases. The experiments are demonstrated using a perdeuterated sample of the chicken α-spectrin SH3 domain.

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Acknowledgments

We thank Nikolai Skrynnikov for many stimulating discussions. This work was supported by the Leibniz-Gemeinschaft and the DFG (grants Re1435, SFB 449, SFB 740).

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Authors and Affiliations

  1. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125, Berlin, Germany

    Veniamin Chevelkov, Uwe Fink & Bernd Reif

  2. Charité Universitätsmedizin, 10115, Berlin, Germany

    Bernd Reif

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  1. Veniamin Chevelkov
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  2. Uwe Fink
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  3. Bernd Reif
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Correspondence to Bernd Reif.

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Chevelkov, V., Fink, U. & Reif, B. Quantitative analysis of backbone motion in proteins using MAS solid-state NMR spectroscopy. J Biomol NMR 45, 197–206 (2009). https://doi.org/10.1007/s10858-009-9348-5

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