Multiple redox and non-redox interactions define 2-Cys peroxiredoxin as a regulatory hub in the chloroplast

Mol Plant. 2009 Nov;2(6):1273-88. doi: 10.1093/mp/ssp089. Epub 2009 Nov 1.

Abstract

In plants, the highly abundant 2-cysteine peroxiredoxin (2-CysPrx) is associated with the chloroplast and involved in protecting photosynthesis. This work addresses the multiple interactions of the 2-CysPrx in the chloroplast, which depend on its redox state. Transcript co-regulation analysis showed a strong linkage to the peptidyl-prolyl-cis/trans isomerase Cyclophilin 20-3 (Cyp20-3) and other components of the photosynthetic apparatus. Co-expression in protoplasts and quantification of fluorescence resonance energy transfer (FRET) efficiency in vivo confirmed protein interactions of 2-CysPrx with Cyp20-3 as well as NADPH-dependent thioredoxin reductase C (NTRC), while thioredoxin x (Trx-x) did not form complexes that could enable FRET. Likewise, changes in FRET of fluorescently labeled 2-CysPrx in vitro and in vivo proved redox dependent dynamics of 2-CysPrx. Addition of Cyp20-3 to an in vitro peroxidase assay with 2-CysPrx had no significant effect on peroxide reduction. Also, in the presence of NTRC, addition of Cyp20-3 did not further enhance peroxide reduction. In addition, 2-CysPrx functioned as chaperone and inhibited aggregation of citrate synthase during heat treatment. This activity was partly inhibited by Cyp20-3. As a new interaction partner of decameric 2-CysPrx, photosystem II could be identified after chloroplast fractionation and in pull-down assays after reconstitution. In summary, the data indicate a dynamic function of plant 2-CysPrx as redox sensor, chaperone, and regulator in the chloroplast with diverse functions beyond its role as thiol peroxidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Chloroplasts / genetics
  • Chloroplasts / physiology*
  • Cysteine / metabolism
  • DNA Primers
  • Fluorescence Resonance Energy Transfer
  • Kinetics
  • Light
  • NADP / metabolism
  • Oxidation-Reduction
  • Oxidative Stress
  • Peroxiredoxins / genetics*
  • Peroxiredoxins / metabolism*
  • Phosphorylation
  • Photosynthesis
  • Photosystem I Protein Complex / genetics
  • Photosystem I Protein Complex / metabolism
  • Transcription, Genetic*

Substances

  • DNA Primers
  • Photosystem I Protein Complex
  • NADP
  • Adenosine Triphosphate
  • Peroxiredoxins
  • Cysteine