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Cryogenic temperature effects and resolution upon slow cooling of protein preparations in solid state NMR

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Abstract

X-ray crystallography using synchrotron radiation and the technique of dynamic nuclear polarization (DNP) in nuclear magnetic resonance (NMR) require samples to be kept at temperatures below 100 K. Protein dynamics are poorly understood below the freezing point of water and down to liquid nitrogen temperatures. Therefore, we investigate the α-spectrin SH3 domain by magic angle spinning (MAS) solid state NMR (ssNMR) at various temperatures while cooling slowly. Cooling down to 95 K, the NMR-signals of SH3 first broaden and at lower temperatures they separate into several peaks. The coalescence temperature differs depending on the individual residue. The broadening is shown to be inhomogeneous by hole-burning experiments. The coalescence behavior of 26 resolved signals (of 62) was compared to water proximity and crystal structure Debye–Waller factors (B-factors). Close proximity to the solvent and large B-factors (i.e. mobility) lead, generally, to a higher coalescence temperature. We interpret a high coalescence temperature as indicative of a large number of magnetically inequivalent populations at cryogenic temperature.

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Acknowledgments

The authors would like to thank Anne Diehl and Kristina Rehbein for preparation of the SH3 sample, and Dr. Rasmus Linser for access to his solvent accessibility calculations. This work was supported by the Deutsche Forschungsgemeinschaft (SFB449), Structural Biology of Membrane Proteins (Award #211800), The European Drug Initiave on Channels and Transports (Award #201924), and Bio-NMR (Award # 261863).

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

    Arne H. Linden, W. Trent Franks, Ümit Akbey, Sascha Lange, Barth-Jan van Rossum & Hartmut Oschkinat

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  1. Arne H. Linden
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  2. W. Trent Franks
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  3. Ümit Akbey
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Correspondence to Hartmut Oschkinat.

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Linden, A.H., Franks, W.T., Akbey, Ü. et al. Cryogenic temperature effects and resolution upon slow cooling of protein preparations in solid state NMR. J Biomol NMR 51, 283–292 (2011). https://doi.org/10.1007/s10858-011-9535-z

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