NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein
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 an...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-03-01
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| Series: | Frontiers in Chemistry |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fchem.2020.00136/full |
| _version_ | 1817988948876591104 |
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| author | Annika Ciragan Sofia M. Backlund Kornelia M. Mikula Hannes M. Beyer O. H. Samuli Ollila Hideo Iwaï |
| author_facet | Annika Ciragan Sofia M. Backlund Kornelia M. Mikula Hannes M. Beyer O. H. Samuli Ollila Hideo Iwaï |
| author_sort | Annika Ciragan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-14T00:41:02Z |
| format | Article |
| id | doaj.art-fa9f4e0518ee43869f0fef3e5dc56c64 |
| institution | Directory Open Access Journal |
| issn | 2296-2646 |
| language | English |
| last_indexed | 2024-04-14T00:41:02Z |
| publishDate | 2020-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Chemistry |
| spelling | doaj.art-fa9f4e0518ee43869f0fef3e5dc56c642022-12-22T02:22:12ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462020-03-01810.3389/fchem.2020.00136512974NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split InteinAnnika CiraganSofia M. BacklundKornelia M. MikulaHannes M. BeyerO. H. Samuli OllilaHideo IwaïThe 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.https://www.frontiersin.org/article/10.3389/fchem.2020.00136/fullinteinsprotein ligationNMR spectroscopysegmental isotopic labelingTonBintrinsically disordered protein |
| spellingShingle | Annika Ciragan Sofia M. Backlund Kornelia M. Mikula Hannes M. Beyer O. H. Samuli Ollila Hideo Iwaï NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein Frontiers in Chemistry inteins protein ligation NMR spectroscopy segmental isotopic labeling TonB intrinsically disordered protein |
| title | NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein |
| title_full | NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein |
| title_fullStr | NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein |
| title_full_unstemmed | NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein |
| title_short | NMR Structure and Dynamics of TonB Investigated by Scar-Less Segmental Isotopic Labeling Using a Salt-Inducible Split Intein |
| title_sort | nmr structure and dynamics of tonb investigated by scar less segmental isotopic labeling using a salt inducible split intein |
| topic | inteins protein ligation NMR spectroscopy segmental isotopic labeling TonB intrinsically disordered protein |
| url | https://www.frontiersin.org/article/10.3389/fchem.2020.00136/full |
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