NirF is a periplasmic protein that binds d1 heme as part of its essential role in d1 heme biogenesis

Bali, Shilpa, Warren, Martin J. ORCID: https://orcid.org/0000-0002-6028-6456 and Ferguson, Stuart J. (2010) NirF is a periplasmic protein that binds d1 heme as part of its essential role in d1 heme biogenesis. FEBS Journal, 277 (23). pp. 4944-4955. ISSN 1742-464X

Full text not available from this repository. (Request a copy)

Abstract

The cytochrome cd1 nitrite reductase from Paracoccus pantotrophus catalyses the one electron reduction of nitrite to nitric oxide using two heme cofactors. The site of nitrite reduction is the d1 heme, which is synthesized under anaerobic conditions by using nirECFD-LGHJN gene products. In vivo studies with an unmarked deletion strain, ΔnirF, showed that this gene is essential for cd1 assembly and consequently for denitrification, which was restored when the ΔnirF strain was complemented with wild-type, plasmid-borne, nirF. Removal of a signal sequence and deletion of a conserved N-terminal Gly-rich motif from the NirF coded on a plasmid resulted in loss of in vivo NirF activity. We demonstrate here that the product of the nirF gene is a periplasmic protein and, hence, must be involved in a late stage of the cofactor biosynthesis. In vitro studies with purified NirF established that it could bind d1 heme. It is concluded that His41 of NirF, which aligns with His200 of the d1 heme domain of cd1, is essential both for this binding and for the production of d1 heme; replacement of His41 by Ala, Cys, Lys and Met all gave nonfunctional proteins. Potential functions of NirF are discussed.

Item Type: Article
Uncontrolled Keywords: cytochrome cd,d heme biosynthesis,denitrification,nitrite reductase,paracoccus pantotrophus,tetrapyrrole,biochemistry,molecular biology,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School: Faculty of Science
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 20 Sep 2022 15:30
Last Modified: 21 Sep 2022 00:23
URI: https://ueaeprints.uea.ac.uk/id/eprint/88521
DOI: 10.1111/j.1742-4658.2010.07899.x

Actions (login required)

View Item View Item