The cysteine-desulfurase IscS promotes the production of the rhodanese RhdA in the persulfurated form

FEBS Lett. 2005 Dec 19;579(30):6786-90. doi: 10.1016/j.febslet.2005.11.013. Epub 2005 Nov 21.

Abstract

After heterologous expression in Escherichia coli, the Azotobacter vinelandii rhodanese RhdA is purified in a persulfurated form (RhdA-SSH). We identified l-cysteine as the most effective sulfur source in producing RhdA-SSH. An E. coli soluble extract was required for in vitro persulfuration of RhdA, and the addition of pyridoxal-5'-phosphate increased RhdA-SSH production, indicating a likely involvement of a cysteine desulfurase. We were able to show the formation of a covalent complex between IscS and RhdA. By combining a time-course fluorescence assay and mass spectrometry analysis, we demonstrated the transfer of sulfur from E. coli IscS to RhdA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azotobacter vinelandii / enzymology
  • Carbon-Sulfur Lyases / chemistry*
  • Carbon-Sulfur Lyases / metabolism*
  • Catalytic Domain
  • Cysteine / chemistry
  • Escherichia coli / genetics
  • Histidine / chemistry
  • Peptide Mapping
  • Protein Structure, Tertiary
  • Spectrometry, Fluorescence
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Sulfhydryl Compounds / chemistry
  • Sulfhydryl Compounds / metabolism
  • Sulfur / chemistry*
  • Sulfur / metabolism
  • Thiosulfate Sulfurtransferase / biosynthesis*
  • Thiosulfate Sulfurtransferase / chemistry
  • Thiosulfate Sulfurtransferase / genetics
  • Thiosulfate Sulfurtransferase / isolation & purification
  • Thiosulfate Sulfurtransferase / metabolism*

Substances

  • Sulfhydryl Compounds
  • Histidine
  • Sulfur
  • Thiosulfate Sulfurtransferase
  • Carbon-Sulfur Lyases
  • cysteine desulfurase
  • Cysteine