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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

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


Authors

Jenny Ni

James L. Wood

Melanie Y. White

Norbert Lihi

Todd E. Markham

Joseph Wang

Rachel Codd



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., …Codd, R. (2023). Reduction-cleavable desferrioxamine B pulldown system enriches Ni( ii )-superoxide dismutase from a Streptomyces proteome. RSC Chemical Biology, 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
DOI https://doi.org/10.1039/d3cb00097d
Public URL https://durham-repository.worktribe.com/output/1797685

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