NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein

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Ciragan , A , Backlund , S M , Mikula , K M , Beyer , H M , Samuli Ollila , O H & Iwaï , H 2020 , ' NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein ' , Frontiers in Chemistry , vol. 8 , 136 . https://doi.org/10.3389/fchem.2020.00136

Title: NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein
Author: Ciragan, Annika; Backlund, Sofia M.; Mikula, Kornelia M.; Beyer, Hannes M.; Samuli Ollila, O. H.; Iwaï, Hideo
Contributor: University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
University of Helsinki, Institute of Biotechnology
Date: 2020-03-19
Number of pages: 13
Belongs to series: Frontiers in Chemistry
ISSN: 2296-2646
URI: http://hdl.handle.net/10138/317514
Abstract: The growing understanding of partially unfolded proteins increasingly points to their biological relevance in allosteric regulation, complex formation, and protein design. However, the structural characterization of disordered proteins remains challenging. NMR methods can access both the dynamics and structures of such proteins, yet suffering from a high degeneracy of NMR signals. Here, we overcame this bottleneck utilizing a salt-inducible split intein to produce segmentally isotope-labeled samples with the native sequence, including the ligation junction. With this technique, we investigated the NMR structure and conformational dynamics of TonB from Helicobacter pylori in the presence of a proline-rich low complexity region. Spin relaxation experiments suggest that the several nano-second time scale dynamics of the C-terminal domain (CTD) is almost independent of the faster pico-to-nanosecond dynamics of the low complexity central region (LCCR). Our results demonstrate the utility of segmental isotopic labeling for proteins with heterogenous dynamics such as TonB and could advance NMR studies of other partially unfolded proteins.
Subject: 116 Chemical sciences
1182 Biochemistry, cell and molecular biology
inteins
protein ligation
NMR spectroscopy
segmental isotopic labeling
TonB
intrinsically disordered protein
multi-domain proteins
protein dynamics
TORSION ANGLE DYNAMICS
C-TERMINAL DOMAIN
BACKBONE DYNAMICS
MULTIDOMAIN PROTEIN
CHEMICAL LIGATION
CRYSTAL-STRUCTURE
ESCHERICHIA-COLI
DNAE INTEIN
SPECTROSCOPY
RELAXATION
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