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Structural heterogeneity in microcrystalline ubiquitin studied by solid‐state NMR
Author(s) -
Fasshuber Hannes Klaus,
Lakomek NilsAlexander,
Habenstein Birgit,
Loquet Antoine,
Shi Chaowei,
Giller Karin,
Wolff Sebastian,
Becker Stefan,
Lange Adam
Publication year - 2015
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2654
Subject(s) - microcrystalline , chemistry , crystallography , solid state nuclear magnetic resonance , solid state , nmr spectra database , carbon 13 nmr , nuclear magnetic resonance spectroscopy , globular protein , conformational isomerism , spectral line , nuclear magnetic resonance , molecule , stereochemistry , physics , organic chemistry , astronomy
By applying [1‐ 13 C]‐ and [2‐ 13 C]‐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.