Isolation and identification of phosphatidic acid targets from plants

Christa Testerink, Henk L. Dekker, Ze Yi Lim, Melloney K. Johns, Andrew B. Holmes, Chris G. De Koster, Nicholas T. Ktistakis, Teun Munnik*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

149 Citations (Scopus)

Abstract

Phosphatidic acid (PA) is emerging as an important lipid signalling molecule. In plants, it is implicated in various stress-signalling pathways and is formed in response to wounding, osmotic stress, cold stress, pathogen elicitors, Nod factors, ethylene and abscisic acid. How PA exerts its effects is still unknown, mainly because of the lack of characterized PA targets. In an approach to isolate such targets we have used PA-affinity chromatography. Several PA-binding proteins were present in the soluble fraction of tomato and Arabidopsis cells. Using mass spectrometric analysis, several of these proteins, including Hsp90, 14-3-3 proteins, an SnRK2 serine/threonine protein kinase and the PP2A regulatory subunit RCN1 could be identified. As an example, the binding of one major PA-binding protein, phosphoenolpyruvate carboxylase (PEPC), was characterized further. Competition experiments with different phospholipids confirmed specificity for PA. Hypo-osmotic treatment of the cells increased the amount of PEPC that bound the PA beads without increasing the absolute amount of PEPC. This suggests that PEPC's affinity for PA had increased. The work shows that PA-affinity chromatography/mass spectrometry is an effective way to isolate and identify PA-binding proteins from plants.

Original languageEnglish
Pages (from-to)527-536
Number of pages10
JournalPlant Journal
Volume39
Issue number4
DOIs
Publication statusPublished - Aug 2004
Externally publishedYes

Keywords

  • Lipid signalling
  • Mass spectrometry
  • Phosphatidic acid
  • Phosphoenolpyruvate carboxylase
  • Plant stress

Fingerprint Dive into the research topics of 'Isolation and identification of phosphatidic acid targets from plants'. Together they form a unique fingerprint.

  • Cite this

    Testerink, C., Dekker, H. L., Lim, Z. Y., Johns, M. K., Holmes, A. B., De Koster, C. G., ... Munnik, T. (2004). Isolation and identification of phosphatidic acid targets from plants. Plant Journal, 39(4), 527-536. https://doi.org/10.1111/j.1365-313X.2004.02152.x