Tools
Tools
Butler, C. S., Charnock, J. M., Garner, C. D., Thomson, A. J., Ferguson, S. J., Berks, B. C. and Richardson, D. J. 
ORCID: https://orcid.org/0000-0002-6847-1832
(2000)
Thiocyanate binding to the molybdenum centre of the periplasmic nitrate reductase from Paracoccus pantotrophus.
Biochemical Journal, 352 (3).
pp. 859-864.
ISSN 0264-6021
Abstract
The periplasmic nitrate reductase (NAP) from Paracoccus pantotrophus is a soluble two-subunit enzyme (NapAB) that binds two haem groups, a [4Fe-4S] cluster and a bis(molybdopterin guanine dinucleotide) (MGD) cofactor that catalyses the reduction of nitrate to nitrite. In the present study the effect of KSCN (potassium thiocyanate) as an inhibitor and Mo ligand has been investigated. Results are presented that show NAP is sensitive to SCN- (thiocyanate) inhibition, with SCN- acting as a competitive inhibitor of nitrate (Ki ≈ 4.0 mM). The formation of a novel EPR Mo(V) species with an elevated gav value (gav ∼ 1.994) compared to the Mo(V) High-g (resting) species was observed upon redox cycling in the presence of SCN-. Mo K-edge EXAFS analysis of the dithionite-reduced NAP was best fitted as a mono-oxo Mo(IV) species with three Mo-S ligands at 2.35 Å (1 Å = 0.1 nm) and a Mo-O ligand at 2.14 Å. The addition of SCN- to the reduced Mo(IV) NAP generated a sample that was best fitted as a mono-oxo (1.70 Å) Mo(IV) species with four Mo-S ligands at 2.34 Å. Taken together, the competitive nature of SCN- inhibition of periplasmic nitrate reductase activity, the elevated Mo(V) EPR gav value following redox cycling in the presence of SCN- and the increase in sulphur co-ordination of Mo(IV) upon SCN- binding, provide strong evidence for the direct binding of SCN- via a sulphur atom to Mo.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | epr spectroscopy,exafs spectroscopy,ligand binding,molybdoenzymes,biochemistry,molecular biology,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1303 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Organisms and the Environment Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 14 Jul 2022 16:31 |
| Last Modified: | 15 May 2023 00:53 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/86123 |
| DOI: | 10.1042/0264-6021:3520859 |
Actions (login required)
 |
View Item |