Including protons in solid-state NMR resonance assignment and secondary structure analysis : the example of RNA polymerase II subunits Rpo4/7/7

  • 1H-detected solid-state NMR experiments feasible at fast magic-angle spinning (MAS) frequencies allow accessing 1H chemical shifts of proteins in solids, which enables their interpretation in terms of secondary structure. Here we present 1H and 13C-detected NMR spectra of the RNA polymerase subunit Rpo7 in complex with unlabeled Rpo4 and use the 13C, 15N, and 1H chemical-shift values deduced from them to study the secondary structure of the protein in comparison to a known crystal structure. We applied the automated resonance assignment approach FLYA including 1H-detected solid-state NMR spectra and show its success in comparison to manual spectral assignment. Our results show that reasonably reliable secondary-structure information can be obtained from 1H secondary chemical shifts (SCS) alone by using the sum of 1Hα and 1HN SCS rather than by TALOS. The confidence, especially at the boundaries of the observed secondary structure elements, is found to increase when evaluating 13C chemical shifts, here either by using TALOS or in terms of 13C SCS.

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Author:Anahit Torosyan, Thomas Wiegand, Maarten Schledorn, Daniel Klose, Peter Güntert, Anja Böckmann, Beat H. Meier
URN:urn:nbn:de:hebis:30:3-514703
DOI:https://doi.org/10.3389/fmolb.2019.00100
ISSN:2296-889X
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/31637245
Parent Title (English):Frontiers in molecular biosciences
Publisher:Frontiers
Place of publication:Lausanne
Contributor(s):Anastasia S. Politou
Document Type:Article
Language:English
Year of Completion:2019
Date of first Publication:2019/10/04
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2019/10/23
Tag:Rpo4/7; carbon and proton assignments; secondary chemical shifts; solid-state NMR; ssFLYA
Volume:6
Issue:Art. 100
Page Number:8
First Page:1
Last Page:8
Note:
Copyright © 2019 Torosyan, Wiegand, Schledorn, Klose, Güntert, Böckmann and Meier. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
HeBIS-PPN:455690634
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Wissenschaftliche Zentren und koordinierte Programme / Zentrum für Biomolekulare Magnetische Resonanz (BMRZ)
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 4.0