Identification of a Nfs1p-bound persulfide intermediate in Fe–S cluster synthesis by intact mitochondria
| Autor: | Heeyong Yoon, Andrew Dancis, Alok Pandey, Debkumar Pain, Elise R. Lyver |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Saccharomyces cerevisiae Sulfurtransferase Sulfides Mitochondrion Sulfur Radioisotopes Aconitase Article Mitochondrial Proteins chemistry.chemical_compound Nucleotide Cysteine Molecular Biology Cysteine metabolism Aconitate Hydratase chemistry.chemical_classification biology Cysteine desulfurase Cell Biology biology.organism_classification Mitochondria chemistry Biochemistry Isotope Labeling Sulfurtransferases Molecular Medicine |
| Zdroj: | Mitochondrion. 12:539-549 |
| ISSN: | 1567-7249 |
| Popis: | 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 [(35)S]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 (1 mM) 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. |
| Databáze: | OpenAIRE |
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