chloroplast NifS-like protein of Arabidopsis thaliana is required for iron-sulfur cluster formation in ferredoxin

Plastids are known to be able to synthesize their own iron—sulfur clusters, but the biochemical machinery responsible for this process is not known. In this study it is investigated whether CpNifS, the chloroplastic NifS-like cysteine desulfurase of Arabidopsis thaliana (L.) Heynh. is responsible fo...

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Bibliographic Details
Published in:Planta 2005-02, Vol.220 (4), p.602-608
Main Authors: Ye, H, Garifullina, G.F, Abdel-Ghany, S.E, Zhang, L, Pilon-Smits, E.A.H, Pilon, M
Format: Article
Language:English
Subjects:
Arabidopsis - metabolism
Arabidopsis Proteins - metabolism
Arabidopsis thaliana
Biological and medical sciences
carbon-sulfur lyases
Chloroplasts
Chloroplasts - metabolism
Chromatography, Gel
cysteine desulfurase
Dimers
enzyme activity
Enzymes
Ferredoxins
Ferredoxins - metabolism
Fundamental and applied biological sciences. Psychology
Genes. Genome
Iron
iron-sulfur cluster
Iron-Sulfur Proteins - metabolism
Kinetics
Microbiology
Mitochondria
Molecular and cellular biology
Molecular genetics
Molecular weight
plant biochemistry
Plants
Plastids
Proteins
Sulfur
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Summary:Plastids are known to be able to synthesize their own iron—sulfur clusters, but the biochemical machinery responsible for this process is not known. In this study it is investigated whether CpNifS, the chloroplastic NifS-like cysteine desulfurase of Arabidopsis thaliana (L.) Heynh. is responsible for the release of sulfur from cysteine for the biogenesis of iron—sulfur (Fe—S) clusters in chloroplasts. Using an in vitro reconstitution assay it was found that purified CpNifS was sufficient for Fe—S cluster formation in ferredoxin in the presence of cysteine and a ferrous iron salt. Antibody-depletion experiments using stromal extract showed that CpNifS is also essential for the Fe—S cluster formation activity of chloroplast stroma. The activity of CpNifS in the stroma was 50- to 80-fold higher than that of purified CpNifS on a per-protein basis, indicating that other stromal factors cooperate in Fe—S cluster formation. When stromal extract was separated on a gel-filtration column, most of the CpNifS eluted as a dimer of 86 kDa, but a minor fraction of the stromal CpNifS eluted at a molecular weight of approx. 600 kDa, suggesting the presence of a multi-protein complex. The possible nature of the interacting proteins is discussed.
ISSN:0032-0935
1432-2048
DOI:10.1007/s00425-004-1388-1