TY - JOUR
T1 - Molecular cloning and functional analysis of polyphosphoinositide- dependent phospholipase D, PLDβ, from Arabidopsis
AU - Pappan, Kirk
AU - Qin, Wensheng
AU - Dyer, James H.
AU - Zheng, Ling
AU - Wang, Xuemin
PY - 1997/3/14
Y1 - 1997/3/14
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0031003039&partnerID=8YFLogxK
U2 - 10.1074/jbc.272.11.7055
DO - 10.1074/jbc.272.11.7055
M3 - Article
C2 - 9054397
AN - SCOPUS:0031003039
SN - 0021-9258
VL - 272
SP - 7055
EP - 7061
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 11
ER -