A novel proteomic approach reveals a role for Mg-protoporphyrin IX in response to oxidative stress

Research output: Contribution to journalArticle

Abstract

The presence of genes encoding organellar proteins in different cellular compartments necessitates a tight coordination of expression by the different genomes of the eukaryotic cell. This coordination of gene expression is achieved by organelle-to-nucleus communication. Stress-induced perturbations of the tetrapyrrole pathway trigger large changes in nuclear gene expression. In order to investigate whether the tetrapyrrole Mg-ProtoIX itself is an important part of plastid-to-nucleus communication, we used an affinity column containing Mg-ProtoIX covalently linked to an Affi-Gel matrix. The proteins that bound to Mg-ProtoIX were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis combined with nano liquid chromatography-mass spectrometry (MS)/MS. Thus, we present a novel proteomic approach to address the mechanisms involved in cellular signaling and we identified interactions between Mg-ProtoIX and a large number of proteins associated with oxidative stress responses. Our approach revealed an interaction between Mg-ProtoIX and the heat shock protein 90-type protein, HSP81-2 suggesting that a regulatory complex including HSP90 proteins and tetrapyrroles controlling gene expression is evolutionarily conserved between yeast and plants. In addition, our list of putative Mg-ProtoIX-binding proteins demonstrated that binding of tetrapyrroles does not depend on a specific amino acid motif but possibly on a specific fold of the protein.

Details

Authors
  • P. Kindgren
  • M. J. Eriksson
  • C. Benedict
  • A. Mohapatra
  • S. P. Gough
  • Mats Hansson
  • T. Kieselbach
  • A. Strand
External organisations
  • Carlsberg Research Center / Carlsberg Laboratory
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology

Keywords

  • Protoporphyrins/*metabolism, Proteomics/*methods, Protein Subunits/metabolism, Protein Binding, *Oxidative Stress, Lyases/metabolism, Light-Harvesting Protein Complexes/metabolism, Plant, Gene Expression Regulation, Computational Biology, Western, Blotting, Arabidopsis Proteins/genetics/metabolism, Amino Acid Motifs, Arabidopsis/enzymology/genetics, Reproducibility of Results, Signal Transduction, Spectrometry, Fluorescence, Stress, Physiological, Tetrapyrroles/metabolism
Original languageEnglish
Pages (from-to)310-320
JournalPhysiologia Plantarum
Volume141
Issue number4
Publication statusPublished - 2011
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes

Bibliographic note

4