Identification of a Nfs1p-bound persulfide intermediate in Fe-S cluster synthesis by intact mitochondria

Alok Pandey, Heeyong Yoon, Elise R. Lyver, Andrew Dancis, Debkumar Pain

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Cysteine desulfurases generate a covalent persulfide intermediate from cysteine, and this activated form of sulfur is essential for the synthesis of iron-sulfur (Fe-S) clusters. In yeast mitochondria, there is a complete machinery for Fe-S cluster synthesis, including a cysteine desulfurase, Nfs1p. Here we show that following supplementation of isolated mitochondria with [35S]cysteine, a radiolabeled persulfide could be detected on Nfs1p. The persulfide persisted under conditions that did not permit Fe-S cluster formation, such as nucleotide and/or iron depletion of mitochondria. By contrast, under permissive conditions, the radiolabeled Nfs1p persulfide was greatly reduced and radiolabeled aconitase was formed, indicating transfer of persulfide to downstream Fe-S cluster recipients. Nfs1p in mitochondria was found to be relatively more resistant to inactivation by N-ethylmaleimide (NEM) as compared with a prokaryotic cysteine desulfurase. Mitochondria treated with NEM (1mM) formed the persulfide on Nfs1p but failed to generate Fe-S clusters on aconitase, likely due to inactivation of downstream recipient(s) of the Nfs1p persulfide. Thus the Nfs1p-bound persulfide as described here represents a precursor en route to Fe-S cluster synthesis in mitochondria.

Original languageEnglish (US)
Pages (from-to)539-549
Number of pages11
JournalMitochondrion
Volume12
Issue number5
DOIs
StatePublished - Sep 2012

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Cell Biology

Keywords

  • Aconitase
  • Cysteine desulfurase
  • Fe-S clusters
  • Nfs1p Isd11p complex
  • Persulfide
  • Yeast

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