Skip to main content
Article thumbnail
Location of Repository

Purification and biochemical characterization of Arabidopsis At-NEET,an ancient iron-sulfur protein, reveals a conserved cleavage motif forsubcellular localization

By Li-Wen Su, Shu Heng Chang, Meng-Ying Li, Heng-Yi Huang, Wann-Neng Jane and Jun-Yi Yang

Abstract

CDGSH iron-sulfur domain-containing proteins (CISDs) are newly discovered proteins with electron-accepting and electron-donating moieties. Although the CISDs of plants and animals show high sequence similarity in their CDGSH domain at the C-terminus, their N-terminal peptides have low sequence homology. Here, we show that At-NEET, a recently identified Arabidopsis CISD, contains a cleavable N-terminal peptide for chloroplast targeting, which is different from the uncleavable N-terminal peptide of mammal CISDs for mitochondrial outer membrane localization. Using affinity purification to isolate endogenous At-NEET, we identified a consensus sequence for the chloroplast transit peptide cleavage site of V-[R/K]↓A-E in At-NEET as well as other plant CISDs. Moreover, chloroplast subfractionation and immunogold labeling experiments showed that At-NEET localizes to the stroma of chloroplast. In addition, biochemical characterization revealed that At-NEET contains a conserved Cys(3)-His(1) ligand in the CDGSH domain, which is essential for coordination of 2Fe-2S clusters and protein folding. Our findings suggest that plant and animal CISDs contain an evolutionarily conserved CDGSH domain. However, they show different subcellular localization patterns that may result in distinct physiological functions

Topics: At-NEET, CDGSH domain, Iron-sulfur protein, Chloroplast transit peptide
Year: 2014
DOI identifier: 10.1016/j.plantsci.2013.09.001
OAI identifier: oai:ir.lib.nchu.edu.tw:11455/85146
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://dx.doi.org/10.1016/j.pl... (external link)
  • http://hdl.handle.net/11455/85... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.