Reduction-cleavable desferrioxamine B pulldown system enriches Ni( ii )-superoxide dismutase from a Streptomyces proteome

Ni, Jenny; Wood, James L.; White, Melanie Y.; Lihi, Norbert; Markham, Todd E.; Wang, Joseph; Chivers, Peter T.; Codd, Rachel

Abstract

Two resins with the hydroxamic acid siderophore desferrioxamine B (DFOB) immobilised as a free ligand or its Fe(iii) complex were prepared to screen the Streptomyces pilosus proteome for proteins involved in siderophore-mediated Fe(iii) uptake. The resin design included a disulfide bond to enable the release of bound proteins under mild reducing conditions. Proteomics analysis of the bound fractions did not identify proteins associated with siderophore-mediated Fe(iii) uptake, but identified nickel superoxide dismutase (NiSOD), which was enriched on the apo-DFOB-resin but not the Fe(iii)-DFOB-resin or the control resin. While DFOB is unable to sequester Fe(iii) from sites deeply buried in metalloproteins, the coordinatively unsaturated Ni(ii) ion in NiSOD is present in a surface-exposed loop region at the N-terminus, which might enable partial chelation. The results were consistent with the notion that the apo-DFOB-resin formed a ternary complex with NiSOD, which was not possible for either the coordinatively saturated Fe(iii)-DFOB-resin or the non-coordinating control resin systems. In support, ESI-TOF-MS measurements from a solution of a model Ni(ii)-SOD peptide and DFOB showed signals that correlated with a ternary Ni(ii)-SOD peptide–DFOB complex. Although any biological implications of a DFOB–NiSOD complex are unclear, the work shows that the metal coordination properties of siderophores might influence an array of metal-dependent biological processes beyond those established in iron uptake.

Citation

Ni, J., Wood, J. L., White, M. Y., Lihi, N., Markham, T. E., Wang, J., Chivers, P. T., & Codd, R. (2023). Reduction-cleavable desferrioxamine B pulldown system enriches Ni( ii )-superoxide dismutase from a Streptomyces proteome. RSC Chemical Biology, 4, Article 1064. https://doi.org/10.1039/d3cb00097d

• Journal Article Type: Article
• Acceptance Date: Oct 2, 2023
• Online Publication Date: Oct 2, 2023
• Publication Date: 2023
• Deposit Date: Oct 16, 2023
• Publicly Available Date: Oct 16, 2023
• Journal: RSC Chemical Biology
• Electronic ISSN: 2633-0679
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 4
• Article Number: 1064
• DOI: https://doi.org/10.1039/d3cb00097d
• Public URL: https://durham-repository.worktribe.com/output/1797685