Molecular cloning and functional analysis of polyphosphoinositide- dependent phospholipase D, PLDβ, from Arabidopsis

Kirk Pappan, Wensheng Qin, Jim Dyer, Ling Zheng, Xuemin Wang

Research output: Contribution to journalArticle

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Abstract

A novel plant phospholipase D (PLD; EC 3.1.4.4) activity, which is dependent on phosphatidylinositol 4,5-bisphosphate (PIP2) and nanomolar concentrations of calcium, has been identified in Arabidopsis. This report describes the cloning, expression, and characterization of an Arabidopsis cDNA that encodes this PLD. We have designated names of PLDβ for this PIP2- dependent PLD and PLDα for the previously characterized PIP2-independent PLD that requires millimolar Ca2+ for optimal activity. The PLDβ cDNA contains an open reading frame of 2904 nucleotides coding for a 968-amino acid protein of 108,575 daltons. Expression of this PLDβ cDNA clone in Escherichia coli results in the accumulation of a functional PLD having PLDβ, but not PLDα, activity. The activity of the expressed PLDβ is dependent on PIP2 and submicromolar amounts of Ca2+, inhibited by neomycin, and stimulated by a soluble factor from plant extracts. Sequence analysis reveals that PLDβ is evolutionarily divergent from PLDα and that its N terminus contains a regulatory Ca2+-dependent phospholipid-binding (C2) domain that is found in a number of signal transducing and membrane trafficking proteins.

Original languageEnglish
Pages (from-to)7055-7061
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number11
DOIs
StatePublished - 14 Mar 1997

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Phosphatidylinositol Phosphates
Phospholipase D
Functional analysis
Cloning
Molecular Cloning
Arabidopsis
Complementary DNA
1-dodecylpyridoxal
Neomycin
Plant Extracts
Phosphatidylinositols
Escherichia coli
Open Reading Frames

Cite this

Pappan, Kirk ; Qin, Wensheng ; Dyer, Jim ; Zheng, Ling ; Wang, Xuemin. / Molecular cloning and functional analysis of polyphosphoinositide- dependent phospholipase D, PLDβ, from Arabidopsis. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 11. pp. 7055-7061.
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Molecular cloning and functional analysis of polyphosphoinositide- dependent phospholipase D, PLDβ, from Arabidopsis. / Pappan, Kirk; Qin, Wensheng; Dyer, Jim; Zheng, Ling; Wang, Xuemin.

In: Journal of Biological Chemistry, Vol. 272, No. 11, 14.03.1997, p. 7055-7061.

Research output: Contribution to journalArticle

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