Stimulation of phosphatidylinositol 4,5-bisphosphate phospholipase C activity by phosphatidic acid

doi:10.1016/0003-9861(89)90318-4

Archives of Biochemistry and Biophysics

Volume 268, Issue 2, 1 February 1989, Pages 516-524

https://doi.org/10.1016/0003-9861(89)90318-4 Get rights and content

Abstract

Phosphatidic acid was a potent activator of the phosphatidylinositol 4,5-bisphosphate (PtdIns-P₂) phospholipase C activity associated with human platelet membranes. Lysophosphatidic acid was half as active as phosphatidic acid, and shortening the fatty acid chain reduced the effectiveness of the corresponding phosphatidic acid. Compounds lacking either the phosphate group (diacylglycerol or phorbol ester) or the fatty acid (glycerol phosphate) were not activators. When the negative charge was contributed by a carboxyl group (fatty acid or phosphatidylserine), stimulation of phospholipase C was weak but detectable. Structural analogs of phosphatidic acid (lipopolysaccharide, lipid A, and 2,3-diacylglucosamine 1-phosphate) were less effective but also enhanced Ptdlnsp₂ hydrolysis. Phosphatidic acid potentiated the activation of phospholipase C by α-thrombin, chelators, and guanine nucleotides. Phosphatidylinositol 4-phosphate and PtdIns-P₂ were also effective activators of PtdIns-P₂ degradation. Other phospholipids were without effect. The production of inositol 1,4,5-trisphosphate and diacylglycerol via the activation of phospholipase C provides a rationale for the cellular responses evoked by phosphatidic acid and the ability of this phospholipid to potentiate and initiate hormonal responses.

References (76)

R.H. Michell
Biochim. Biophys. Acta
(1975)
C. Serhan et al.
J. Biol. Chem
(1981)
C. Serhan et al.
J. Biol. Chem
(1982)
R. Nayar et al.
Biochim. Biophys. Acta
(1984)
E.G. Lapetina et al.
J. Biol. Chem
(1981)
A.M. Benton et al.
Blood
(1982)
S.P. Watson et al.
Biochem. Biophys. Res. Commun
(1985)
M.-F. Simon et al.
FEBS Lett
(1984)
J.N. Fain
Vitam. Horm
(1984)
T. Murayama et al.
J. Biol. Chem
(1987)

Cited by (80)

Phospholipase C and D regulation of Src, calcium release and membrane fusion during Xenopus laevis development
2015, Developmental Biology
Citation Excerpt :
PA (but typically not PS, PC, LPA or PE) stimulated the in vitro activity of PLC from Xenopus oocytes (Jacob et al., 1993), rat (Zhou et al., 1999), bacteria (Wehbi et al., 2003), rabbit platelets (Hashizume et al., 1992), human renal cells (Knauss et al., 1990), myocytes (Kurz et al., 1993; Liu et al., 1999), neutrophils (Siddiqui and English, 2000), bovine parathyroid cells (McGhee and Shoback, 1990), 3T3 cells (Murayama and Ui, 1987), snaptosome membranes (Qian and Drewes, 1991), and human platelets (Jackowski and Rock, 1989). In the latter study by experienced lipid biochemists, Jackowski and Rock (1989) showed that PA stimulated soluble or membrane associated PLC activity 3-fold, changed PLC kinetics from rectangular hyperbola to sigmoid, decreased Km but not Vm, and the activation was independent of chelation of Ca2+. They also suggest that PA does not affect substrate PI45P2 availability but acts by binding to and inducing an activating conformational change in PLC itself.
This review emphasizes how lipids regulate membrane fusion and the proteins involved in three developmental stages: oocyte maturation to the fertilizable egg, fertilization and during first cleavage. Decades of work show that phosphatidic acid (PA) releases intracellular calcium, and recent work shows that the lipid can activate Src tyrosine kinase or phospholipase C during Xenopus fertilization. Numerous reports are summarized to show three levels of increase in lipid second messengers inositol 1,4,5-trisphosphate and sn 1,2-diacylglycerol (DAG) during the three different developmental stages. In addition, possible roles for PA, ceramide, lysophosphatidylcholine, plasmalogens, phosphatidylinositol 4-phosphate, phosphatidylinositol 5-phosphate, phosphatidylinositol 4,5-bisphosphate, membrane microdomains (rafts) and phosphatidylinositol 3,4,5-trisphosphate in regulation of membrane fusion (acrosome reaction, sperm–egg fusion, cortical granule exocytosis), inositol 1,4,5-trisphosphate receptors, and calcium release are discussed. The role of six lipases involved in generating putative lipid second messengers during fertilization is also discussed: phospholipase D, autotaxin, lipin1, sphingomyelinase, phospholipase C, and phospholipase A2. More specifically, proteins involved in developmental events and their regulation through lipid binding to SH3, SH4, PH, PX, or C2 protein domains is emphasized. New models are presented for PA activation of Src (through SH3, SH4 and a unique domain), that this may be why the SH2 domain of PLCγ is not required for Xenopus fertilization, PA activation of phospholipase C, a role for PA during the calcium wave after fertilization, and that calcium/calmodulin may be responsible for the loss of Src from rafts after fertilization. Also discussed is that the large DAG increase during fertilization derives from phospholipase D production of PA and lipin dephosphorylation to DAG.
Tight binding inhibition of protein phosphatase-1 by phosphatidic acid. Specificity of inhibition by the phospholipid
2002, Journal of Biological Chemistry
Phosphatidic acid (PA) has been identified as a bioactive lipid second messenger, yet despite extensive investigation, no cellular target has emerged as a mediator of its described biological effects. In this study, we identify the γ isoform of the human protein phosphatase-1 catalytic subunit (PP1cγ) as a high affinity in vitro target of PA. PA inhibited the enzyme dose-dependently with an IC₅₀ of 15 nm. Mechanistically, PA inhibited the enzyme noncompetitively with the kinetics of a tight binding inhibitor and aK_i value of 0.97 ± 0.24 nm. Together, these data describe one of the most potent in vitro effects of PA. To further elucidate the interaction between PA and PP1cγ, structure/function analysis of the lipid was carried out using commercially available and synthetically generated analogs of PA. These studies disclosed that the lipid-protein interaction is dependent on the presence of the lipid phosphate as well as the presence of the fatty acid side chains, because lipids lacking either of these substituents resulted in complete loss of inhibition. However, the specific composition of the fatty acid side chains was not important for inhibition. Using 1-O-hexadecyl,2-oleoyl-PA, it was also shown that the carbonyl group of the sn-1 acyl linkage is not required for the lipid-protein interaction. Finally, using a lipid-protein overlay assay, it was demonstrated that PP1cγ specifically and directly interacts with phosphatidic acid while not significantly binding other phospholipids. These results identify PA as a tight binding and specific inhibitor of PP1, and they raise the hypothesis that PP1cγ may function as a mediator of PA action in cells. They also argue for the existence of a specific high affinity PA-binding domain on the enzyme.
Phospholipase D and immune receptor signalling
2002, Seminars in Immunology
Immune receptors are coupled to the activation of phosphatidylcholine phospholipase D (PC-PLD) that hydrolyses phosphatidylcholine to generate phosphatidic acid and choline. As these receptors are also coupled to other signalling cascades, it has been difficult to define the precise cell activation events resulting from PLD activation in the absence of specific inhibitors. There is increasing evidence that phosphatidic acid acts as an intracellular signalling molecule regulating release of calcium from intracellular stores, sphingosine kinase and protein kinase C activation and membrane budding. Phosphatidic acid can also be rapidly converted into lysophosphatidic acid, diacylglycerol and arachidonates.
Functional coupling of FcγRI to nicotinamide adenine dinucleotide phosphate (reduced form) oxidative burst and immune complex trafficking requires the activation of phospholipase D1
2001, Blood
Citation Excerpt :
The immediate products of PLD hydrolysis of phosphatidylcholine are phosphatidic acid and choline.8 A role for phosphatidic acid as a key intracellular signaling molecule has been proposed as it has been shown to directly activate protein kinases,18,19,26,27 protein tyrosine phosphatase,28-30 phospholipase C,31phosphoinositol-4-kinase,32 sphingosine kinase,33 and small molecular weight guanosine triphosphatase–activating proteins.34 Phosphatidic acid has also been shown to promote the release of calcium from intracellular compartments35,36 and, in neutrophils, to activate the oxidative burst through nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase.24,27
Immunoglobulin G (IgG) receptors (FcγRs) on myeloid cells are responsible for the internalization of immune complexes. Activation of the oxidase burst is an important component of the integrated cellular response mediated by Fc receptors. Previous work has demonstrated that, in interferon-γ–primed U937 cells, the high-affinity receptor for IgG, FcγRI, is coupled to a novel intracellular signaling pathway that involves the sequential activation of phospholipase D (PLD), sphingosine kinase, and calcium transients. Here, it is shown that both known PLD isozymes, PLD1 and PLD2, were present in these cells. With the use of antisense oligonucleotides to specifically reduce the expression of either isozyme, PLD1, but not PLD2, was found to be coupled to FcγRI activation and be required to mediate receptor activation of sphingosine kinase and calcium transients. In addition, coupling of FcγRI to activation of the nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase burst was inhibited by pretreating cells with 0.3% butan-1-ol, indicating an absolute requirement for PLD. Furthermore, use of antisense oligonucleotides to reduce expression of PLD1 or PLD2 demonstrated that PLD1 is required to couple FcγRI to the activation of NADPH oxidase and trafficking of internalized immune complexes for degradation. These studies demonstrate the critical role of PLD1 in the intracellular signaling cascades initiated by FcγRI and its functional role in coordinating the response to antigen-antibody complexes.
Activation of phospholipase C δ1 through C2 domain by a Ca<sup>2+</sup>-enzyme- phosphatidylserine ternary complex
1999, Journal of Biological Chemistry
The concentration of free Ca²⁺and the composition of nonsubstrate phospholipids profoundly affect the activity of phospholipase C δ1 (PLCδ1). The rate of PLCδ1 hydrolysis of phosphatidylinositol 4,5-bisphosphate was stimulated 20-fold by phosphatidylserine (PS), 4-fold by phosphatidic acid (PA), and not at all by phosphatidylethanolamine or phosphatidylcholine (PC). PS reduced the Ca²⁺ concentration required for half-maximal activation of PLCδ1 from 5.4 to 0.5 μm. In the presence of Ca²⁺, PLCδ1 specifically bound to PS/PC but not to PA/PC vesicles in a dose-dependent and saturable manner. Ca²⁺ also bound to PLCδ1 and required the presence of PS/PC vesicles but not PA/PC vesicles. The free Ca²⁺concentration required for half-maximal Ca²⁺ binding was estimated to be 8 μm. Surface dilution kinetic analysis revealed that the K_m was reduced 20-fold by the presence of 25 mol % PS, whereas V_max and K_d were unaffected. Deletion of amino acid residues 646–654 from the C2 domain of PLCδ1 impaired Ca²⁺binding and reduced its stimulation and binding by PS. Taken together, the results suggest that the formation of an enzyme-Ca²⁺-PS ternary complex through the C2 domain increases the affinity for substrate and consequently leads to enzyme activation.
A novel protein kinase target for the lipid second messenger phosphatidic acid
1999, Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Activation of phospholipase D occurs in response to a wide variety of hormones, growth factors, and other extracellular signals. The initial product of phospholipase D, phosphatidic acid (PA), is thought to serve a signaling function, but the intracellular targets for this lipid second messenger are not clearly identified. The production of PA in human neutrophils is closely correlated with the activation of NADPH oxidase, the enzyme responsible for the respiratory burst. We have developed a cell-free system, in which the activation of NADPH oxidase is induced by the addition of PA. Characterization of this system revealed that a multi-functional cytosolic protein kinase was a target for PA, and that two NADPH oxidase components were substrates for the enzyme. Partial purification of the PA-activated protein kinase separated the enzyme from known protein kinase targets of PA. The partially purified enzyme was selectively activated by PA, compared to other phospholipids, and phosphorylated the oxidase component p47-phox on both serine and tyrosine residues. PA-activated protein kinase activity was present in a variety of hematopoietic cells and cell lines and in rat brain, suggesting it has widespread distribution. We conclude that this protein kinase may be a novel target for the second messenger function of PA.

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^☆: This work was supported by National Institutes of Health Grant HL 40560, American Cancer Society Grant BC-582, Cancer Center (CORE) Support Grant CA21765, and the American Lebanese Syrian Associated Charities.

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Stimulation of phosphatidylinositol 4,5-bisphosphate phospholipase C activity by phosphatidic acid☆

Abstract

Biochim. Biophys. Acta

J. Biol. Chem

J. Biol. Chem

Biochim. Biophys. Acta

J. Biol. Chem

Blood

Biochem. Biophys. Res. Commun

FEBS Lett

Vitam. Horm

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

Anal. Biochem

J. Biol. Chem

J. Biol. Chem

Biochim. Biophys. Acta

FEBS Lett

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Mol. Biol

J. Biol. Chem

J. Biol. Chem

Cell. Immunol

Cell

Blood

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem

Biochim. Biophys. Acta

Cell

Cell

Biochem. Biophys. Res. Commun