Plant PIP2-dependent phospholipase D activity is regulated by phosphorylation

FEBS Lett. 2003 Nov 6;554(1-2):50-4. doi: 10.1016/s0014-5793(03)01093-7.


Phospholipase D (PLD) forms the major family of phospholipases that was first discovered and cloned in plants. In this report we have shown, for the first time, that C2 phosphatidylinositol-4,5-bisphosphate (PIP2)-dependent PLD(s) from 5 day hypocotyls of Brassica oleracea associated with plasma membrane is covalently modified-phosphorylated. Pre-incubation of the plasma membrane fraction with acid phosphatase resulted in concentration-dependent inhibition of PIP2-dependent PLD activity. Using matrix-assisted laser desorption/ionization time of flight mass spectrometry of tryptic in-gel digests, the BoPLDgamma(1,2) isoform was identified. Comparing the spectra of the proteins obtained from the plasma membrane fractions treated and non-treated with acid phosphatase, three peptides differing in the mass of the phosphate group (80 Da) were revealed: TMQMMYQTIYK, EVADGTVSVYNSPR and KASKSRGLGK which possess five potential Ser/Thr phosphorylation sites. Our findings suggest that a phosphorylation/dephosphorylation mechanism may be involved in the regulation of plant PIP2-dependent PLDgamma activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acid Phosphatase / metabolism
  • Amino Acid Sequence
  • Binding Sites
  • Brassica / enzymology
  • Membrane Proteins / metabolism
  • Peptide Fragments
  • Phosphatidylinositol 4,5-Diphosphate / metabolism*
  • Phospholipase D / metabolism*
  • Phosphorylation
  • Plant Proteins / metabolism
  • Sequence Alignment
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization


  • Membrane Proteins
  • Peptide Fragments
  • Phosphatidylinositol 4,5-Diphosphate
  • Plant Proteins
  • Acid Phosphatase
  • Phospholipase D