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Plant PIP2-dependent phospholipase D activity is regulated by phosphorylation
Zuzana Novotná1, Jan Linek, Radovan Hynek
1Department of Biochemistry and Microbiology, Institute of Chemical Technology Prague, Prague 6, Czech Republic. zuzana.novotna@vscht.cz
FEBS Letters
|November 5, 2003
Summary
This study reveals that plant Phospholipase D (PLD) is phosphorylated, suggesting a regulatory mechanism. This phosphorylation impacts the activity of phosphatidylinositol-4,5-bisphosphate (PIP2)-dependent PLD in Brassica oleracea.
Area of Science:
- Plant biochemistry
- Enzymology
- Molecular biology
Background:
- Phospholipase D (PLD) is a major enzyme family, initially identified in plants.
- Plant PLDs play crucial roles in cellular signaling and membrane trafficking.
Purpose of the Study:
- To investigate the post-translational modification of phosphatidylinositol-4,5-bisphosphate (PIP2)-dependent Phospholipase D (PLD) in Brassica oleracea.
- To elucidate the role of phosphorylation in regulating plant PLD activity.
Main Methods:
- Isolation and fractionation of plasma membrane from Brassica oleracea hypocotyls.
- Enzymatic assays to measure PLD activity following acid phosphatase treatment.
- Proteomic analysis using MALDI-TOF mass spectrometry to identify protein modifications.
Main Results:
- C2 phosphatidylinositol-4,5-bisphosphate (PIP2)-dependent PLD associated with the plasma membrane is covalently phosphorylated.
- Acid phosphatase treatment inhibited PIP2-dependent PLD activity in a concentration-dependent manner.
- Mass spectrometry identified the BoPLDgamma(1,2) isoform and revealed phosphorylation on specific peptides.
Conclusions:
- Phosphorylation/dephosphorylation likely regulates the activity of plant PIP2-dependent PLDgamma.
- This finding provides novel insights into the molecular mechanisms controlling plant PLD function.