Insights into periplasmic nitrate reductase function under single turnover.

Autor: McGarry J; Department of Chemistry and Biochemistry, University of Wisconsin- Milwaukee, Milwaukee, WI, 53211, USA., Mintmier B; Department of Chemistry and Chemical Biology, Indiana University Indianapolis, Indianapolis, IN, 46202, USA., Metzger MC; Department of Chemistry and Chemical Biology, Indiana University Indianapolis, Indianapolis, IN, 46202, USA., Giri NC; Department of Chemistry and Chemical Biology, Indiana University Indianapolis, Indianapolis, IN, 46202, USA., Britt N; Department of Chemistry and Biochemistry, University of Wisconsin- Milwaukee, Milwaukee, WI, 53211, USA., Basu P; Department of Chemistry and Chemical Biology, Indiana University Indianapolis, Indianapolis, IN, 46202, USA. Basup@iu.edu., Wilcoxen J; Department of Chemistry and Biochemistry, University of Wisconsin- Milwaukee, Milwaukee, WI, 53211, USA. Jarettw@uwm.edu.
Jazyk: angličtina
Zdroj: Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry [J Biol Inorg Chem] 2024 Dec; Vol. 29 (7-8), pp. 811-819. Date of Electronic Publication: 2024 Dec 04.
DOI: 10.1007/s00775-024-02087-5
Abstrakt: Nitrate reductases play pivotal roles in nitrogen metabolism by leveraging the molybdopterin cofactor to facilitate the reduction of nitrate to nitrite. Periplasmic nitrate reductases (NapA) utilize nitrate as a terminal electron acceptor when oxygen is limiting, helping to drive anaerobic metabolism in bacteria. Despite extensive research into NapA homologs, open questions about the mechanism remain especially at the molecular level. More broadly, little is understood of how the molybdopterin cofactor is tuned for catalysis in these enzymes enabling broad substrate scope and reactivity observed in molybdenum-containing enzymes. Here, we have prepared NapA from Campylobacter jejuni under single turnover conditions to generate a singly reduced enzyme that can be further examined by electron paramagnetic resonance (EPR) spectroscopy. Our results provide new context into the known spectra and related structures of NapA and related enzymes. These insights open new avenues for understanding nitrate reductase mechanisms, molybdenum coordination dynamics, and the role of pyranopterin ligands in catalysis.
Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests. Ethical approval: Not applicable.
(© 2024. The Author(s), under exclusive licence to Society for Biological Inorganic Chemistry (SBIC).)
Databáze: MEDLINE
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