Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Jun 27;399(7):787-798.
doi: 10.1515/hsz-2018-0142.

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

Lisa M Galle  1 , George E Cutsail Iii  2 , Volker Nischwitz  3 , Serena DeBeer  2 , Ingrid Span  1
Affiliations

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

Lisa M Galle et al. Biol Chem. .

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.

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

PubMed Disclaimer

References

    1. Anglin, J.R. and Davison, A. (1975). Iron(II) and cobalt(II) complexes of Boc-(Gly-L-Cys-Gly)4-NH2 as analogs for the active site of the iron-sulfur protein rubredoxin. Inorg. Chem. 14, 234–237.
    1. Bencini, A., Bertini, I., Canti, G., Gatteschi, D., and Luchinat, C. (1981). The epr spectra of the inhibitor derivatives of cobalt carbonic anhydrase. J. Inorg. Biochem. 14, 81–93.
    1. Bordeaux, M., Galarneau, A., and Drone, J. (2012). Catalytic, mild, and selective oxyfunctionalization of linear alkanes: current challenges. Angew. Chem. Int. Ed. 51, 10712–10723.
    1. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.
    1. Castillo, R.G., Banerjee, R., Allpress, C.J., Rohde, G.T., Bill, E., Que, L., Lipscomb, J.D., and DeBeer, S. (2017). High-energy-resolution fluorescence-detected X-ray absorption of the Q intermediate of soluble methane monooxygenase. J. Am. Chem. Soc. 139, 18024–18033.

Publication types

LinkOut - more resources