Structural heterogeneity in microcrystalline ubiquitin studied by solid-state NMR |
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| Authors: | Hannes Klaus Fasshuber Nils-Alexander Lakomek Birgit Habenstein Antoine Loquet Chaowei Shi Karin Giller Sebastian Wolff Stefan Becker Adam Lange |
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| Affiliation: | 1. Department of NMR‐Based Structural Biology, Max Planck Institute for Biophysical Chemistry, G?ttingen, Germany;2. Leibniz‐Institut für Molekulare Pharmakologie, Berlin, Germany;3. Institut für Biologie, Humboldt‐Universit?t, zu Berlin, Berlin, Germany;4. Department of Chemistry and Applied Biosciences, ETH Zürich, Zürich, Switzerland;5. Institut Européen de Chimie et de Biologie, Université de Bordeaux CBMN, Pessac, France |
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| Abstract: | By applying [1-13C]- and [2-13C]-glucose labeling schemes to the folded globular protein ubiquitin, a strong reduction of spectral crowding and increase in resolution in solid-state NMR (ssNMR) spectra could be achieved. This allowed spectral resonance assignment in a straightforward manner and the collection of a wealth of long-range distance information. A high precision solid-state NMR structure of microcrystalline ubiquitin was calculated with a backbone rmsd of 1.57 to the X-ray structure and 1.32 Å to the solution NMR structure. Interestingly, we can resolve structural heterogeneity as the presence of three slightly different conformations. Structural heterogeneity is most significant for the loop region β1-β2 but also for β-strands β1, β2, β3, and β5 as well as for the loop connecting α1 and β3. This structural polymorphism observed in the solid-state NMR spectra coincides with regions that showed dynamics in solution NMR experiments on different timescales. |
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| Keywords: | ubiquitin 13C sparse labeling heterogeneity dynamics solid-state NMR |
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