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Paramagnetic Solid-State NMR to Localize the Metal-Ion Cofactor in an Oligomeric DnaB Helicase.
- Source :
-
Chemistry (Weinheim an der Bergstrasse, Germany) [Chemistry] 2021 May 17; Vol. 27 (28), pp. 7745-7755. Date of Electronic Publication: 2021 May 02. - Publication Year :
- 2021
-
Abstract
- Paramagnetic metal ions can be inserted into ATP-fueled motor proteins by exchanging the diamagnetic Mg <superscript>2+</superscript> cofactor with Mn <superscript>2+</superscript> or Co <superscript>2+</superscript> . Then, paramagnetic relaxation enhancement (PRE) or pseudo-contact shifts (PCSs) can be measured to report on the localization of the metal ion within the protein. We determine the metal position in the oligomeric bacterial DnaB helicase from Helicobacter pylori complexed with the transition-state ATP-analogue ADP:AlF <subscript>4</subscript> <superscript>-</superscript> and single-stranded DNA using solid-state NMR and a structure-calculation protocol employing CYANA. We discuss and compare the use of Mn <superscript>2+</superscript> and Co <superscript>2+</superscript> in localizing the ATP cofactor in large oligomeric protein assemblies. <superscript>31</superscript> P PCSs induced in the Co <superscript>2+</superscript> -containing sample are then used to localize the DNA phosphate groups on the Co <superscript>2+</superscript> PCS tensor surface enabling structural insights into DNA binding to the DnaB helicase.<br /> (© 2021 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)
Details
- Language :
- English
- ISSN :
- 1521-3765
- Volume :
- 27
- Issue :
- 28
- Database :
- MEDLINE
- Journal :
- Chemistry (Weinheim an der Bergstrasse, Germany)
- Publication Type :
- Academic Journal
- Accession number :
- 33822417
- Full Text :
- https://doi.org/10.1002/chem.202100462