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

Ni, Jenny and Wood, James L. and White, Melanie Y. and Lihi, Norbert and Markham, Todd E. and Wang, Joseph and Chivers, Peter T. and Codd, Rachel (2023) Reduction-cleavable desferrioxamine B pulldown system enriches Ni(II)-superoxide dismutase from a Streptomyces proteome. RSC CHEMICAL BIOLOGY, 4 (12). pp. 1064-1072. ISSN 2633-0679

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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 sidero- phores might influence an array of metal-dependent biological processes beyond those established in iron uptake.

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