Regulation of phospholipase C-β1 activity by phosphatidic acid

Irene Litosch

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The role of phosphatidic acid (PA) in regulating phospholipase C-β1 (PLC-β1) activity was determined. PA promoted the binding of PLC-β1 to sucrose-loaded unilamellar vesicles (SLUV) containing phosphatidylcholine. PA increased enzymatic activity over a range of Ca2+ concentrations and reduced the Ca2+ concentration required for half-maximal stimulation of activity. PA did not affect the apparent K(m) for phosphatidylinositol 4,5- bisphosphate. Lysophosphatidic acid also enhanced the binding of PLCβ1 to SLUV but was less effective in stimulating enzymatic activity. Diacylglycerol, phosphatidylserine, and oleic acid had little effect on activity. Anionic and neutral detergents did not stimulate activity. PA stimulation was relatively independent of acyl chain length. Dipalmitoyl-PA (16:0) was comparable to PA from egg lecithin and dimyristoyl-PA (C14:0) in stimulating activity, while dilauroyl-PA (C12:0) was slightly less effective. A 100 kDa catalytic fragment of PLC-β1 lacking amino acid residues C- terminal to His880 did not bind to PA and was insensitive to stimulation by 7-15 mol % PA. Stimulation of 100 kDa enzymatic activity required 30 mol % PA. PA increased receptor-G protein stimulation of PLC-β1 activity in membranes. These results demonstrate that PA stimulates basal and receptor-G protein-regulated PLC-β1 activity. PA stimulation occurs through both a C- terminal-dependent and an independent mechanism. The C-terminal-mediated mechanism for stimulation may constitute an important pathway for conferring specific regulation of PLC-β1 in response to increases in cellular PA levels.

Original languageEnglish
Pages (from-to)7736-7743
Number of pages8
JournalBiochemistry
Volume39
Issue number26
DOIs
StatePublished - Jul 4 2000
Externally publishedYes

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Phosphatidic Acids
Type C Phospholipases
Unilamellar Liposomes
GTP-Binding Proteins
Sucrose
Lecithins
Phosphatidylserines
Diglycerides
Oleic Acid
Phosphatidylinositols
Phosphatidylcholines
Chain length
Detergents

ASJC Scopus subject areas

  • Biochemistry

Cite this

Regulation of phospholipase C-β1 activity by phosphatidic acid. / Litosch, Irene.

In: Biochemistry, Vol. 39, No. 26, 04.07.2000, p. 7736-7743.

Research output: Contribution to journalArticle

Litosch, Irene. / Regulation of phospholipase C-β1 activity by phosphatidic acid. In: Biochemistry. 2000 ; Vol. 39, No. 26. pp. 7736-7743.
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