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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred


IMPACT FACTOR 2018: 3.014
5-year IMPACT FACTOR: 3.162

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1437-4315
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Volume 399, Issue 7

Issues

Spectroscopic characterization of the Co-substituted C-terminal domain of rubredoxin-2

Lisa M. Galle
  • Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstr. 1, D-40225 Düsseldorf, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ George E. Cutsail III / Volker Nischwitz
  • Central Institute for Engineering, Electronics and Analytics (ZEA-3), Forschungszentrum Jülich, D-52425 Jülich, Germany
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Serena DeBeer / Ingrid SpanORCID iD: http://orcid.org/0000-0002-2892-4825
Published Online: 2018-06-12 | DOI: https://doi.org/10.1515/hsz-2018-0142

Abstract

Pseudomonas putida rubredoxin-2 (Rxn2) is an essential member of the alkane hydroxylation pathway and transfers electrons from a reductase to the membrane-bound hydroxylase. The regioselective hydroxylation of linear alkanes is a challenging chemical transformation of great interest for the chemical industry. Herein, we report the preparation and spectroscopic characterization of cobalt-substituted P. putida Rxn2 and a truncated version of the protein consisting of the C-terminal domain of the protein. Our spectroscopic data on the Co-substituted C-terminal domain supports a high-spin Co(II) with a distorted tetrahedral coordination environment. Investigation of the two-domain protein Rxn2 provides insights into the metal-binding properties of the N-terminal domain, the role of which is not well understood so far. Circular dichroism, electron paramagnetic resonance and X-ray absorption spectroscopies support an alternative Co-binding site within the N-terminal domain, which appears to not be relevant in nature. We have shown that chemical reconstitution in the presence of Co leads to incorporation of Co(II) into the active site of the C-terminal domain, but not the N-terminal domain of Rxn2 indicating distinct roles for the two rubredoxin domains.

This article offers supplementary material which is provided at the end of the article.

Keywords: AlkG; iron-sulfur protein; metal substitution; Pseudomonas putida GPo1; rubredoxin

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About the article

Received: 2018-01-31

Accepted: 2018-04-25

Published Online: 2018-06-12

Published in Print: 2018-06-27


Conflict of interest statement: The authors declare that they have no conflict of interest regarding the contents of this article.


Citation Information: Biological Chemistry, Volume 399, Issue 7, Pages 787–798, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/hsz-2018-0142.

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