Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.4.3 (phospholipase C)
 18,461 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The beta and gamma subunits of the heterotrimeric guanine nucleotide binding (G protein) act as a dimer and directly regulate various signal transduction pathways. By using cotransfection assays, we tested the ability of several beta gamma combinations to activate inositol phospholipid-specific phospholipase C (PI-PLC)-beta 2. Our findings indicate that only beta gamma combinations that form dimers will activate PI-PLC-beta 2. Since G beta 1 interacts with G gamma 1, while G beta 2 cannot, chimeras between G beta 1 and G beta 2 were used to identify the regions in beta 1 that determine its specific association with gamma 1. Our evidence demonstrates that a chimera between beta 2 and beta 1 that contains the C-terminal 173 amino acids of beta 1 can interact and activate PI-PLC-beta 2 with gamma 1. Chimeras that contain portions of the beta 1 C-terminal region display a weaker association with gamma 1. Furthermore, the contribution of each of these regions depends on the sequence context of each chimeric protein. However, the segment between residues 210 and 293 of beta 1 consistently plays a critical role in specifying association with gamma 1.
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PMID:A segment of the C-terminal half of the G-protein beta 1 subunit specifies its interaction with the gamma 1 subunit. 789 15

The levels of expression of phosphoinositide-specific phospholipase Cs (PLCs) were examined in a series of primary human colon carcinomas and in eight colon carcinoma cell lines by using monoclonal antibodies and cDNA probes for PLC gamma 1, PLC beta 1, and PLC delta 1. Western and northern blot analyses of PLC gamma 1 revealed elevated expression of this isozyme at both the protein and mRNA levels in most tumors when compared with paired adjacent normal mucosa samples (in 11 of 13 pairs in the western blots and 8 of 9 pairs in the northern blots). On the other hand, decreased levels of the PLC delta 1 protein were seen in most colon carcinomas (12 of 13 paired samples). The levels of PLC beta 1 protein were too low to detect possible differences between the carcinoma and normal mucosa samples. Relatively high expression of PLC gamma 1 was found in almost all of the eight human colon carcinoma cell lines at both the protein and mRNA levels. Only weak expression of PLC beta 1 was detected in these cell lines, by both western and northern blot analyses, and PLC delta 1 protein was not detected in any of the carcinoma cell lines. These findings provide evidence that colon carcinomas display altered expression of individual isoforms of PLCs and suggest that increased expression of PLC gamma 1 may play an important role in colon carcinogenesis.
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PMID:Expression of phospholipases gamma 1, beta 1, and delta 1 in primary human colon carcinomas and colon carcinoma cell lines. 789 68

We previously demonstrated that antidepressant drugs (ADs) cause Ca2+ release from inositol 1,4,5-trisphosphate-sensitive Ca2+ stores in cultured neurons of rat frontal cortex. The present study examines the mechanism by which tricyclic ADs activate phospholipase C (PLC) in rat frontal cortex. Using an exogenous substrate to measure PLC activity, we demonstrated that a tricyclic AD, imipramine, stimulated PLC activity of the frontal cortex membrane in a concentration-dependent manner. Two tricyclic ADs, desipramine and amitriptyline, also stimulated PLC activity, while Li+ or pargyline had no effect on PLC activity. Although imipramine did not activate PLC in the membrane in the absence of Ca2+, imipramine synergistically activated PLC in the presence of Ca2+. This result indicates that the mechanism of PLC activation by imipramine is different from its activation by Ca2+. Imipramine stimulated PLC activity in the cytosol of rat frontal cortex as well as in the membrane. Preincubation of the cytosol with anti-PLC-beta 1 antibody prevented the imipramine-mediated activation of PLC. However, preincubation with anti-PLC-gamma 1 or anti-PLC-delta 1 did not prevent activation of PLC. These results suggest that imipramine activates PLC-beta 1 directly without receptor or guanine nucleotide binding protein mediation.
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PMID:Imipramine stimulates phospholipase C activity in rat brain. 789 33

The fourth member of mammalian beta-type phospholipase C isozymes, PLC-beta 4, was recently purified from bovine retina, and the corresponding cDNA was cloned from rat brain and sequenced. PLC-beta 4 has now been shown to differ from the other three mammalian beta-type isozymes (PLC-beta 1, -beta 2, and -beta 3) in that it is selectively inhibited by ribonucleotides. The inhibition requires the 5'-phosphate and 2'-hydroxyl groups of ribose as well as the base moiety. Thus, deoxyribonucleotides and ribose 5-phosphate were not inhibitory. The monophosphate, diphosphate, and triphosphate nucleoside derivatives were all inhibitory, whereas cyclic nucleotides were ineffective. Purine nucleotides were more potent inhibitors than pyrimidine nucleotides; the 50% inhibitory concentrations were 20-30 microM for AMP and GMP, and 100-200 microM for UMP and CMP. Unlike the other beta-type isozymes, PLC-beta 4 contains the GX4GKS consensus sequence for the recognition of the phosphoryl group of nucleotides. In the absence of ribonucleotides, the specific activity of PLC-beta 4 toward phosphatidyl-inositol 4,5-bisphosphate was four to five times the average specific activity of PLC-beta 1 and PLC-beta 3. Thus, nucleotide-dependent inhibition may serve to reduce the activity of PLC-beta 4 in the absence of a hormonal signal. The regulation of PLC-beta 4 by G-proteins was also studied. Similar to the other three PLC-beta isozymes, PLC-beta 4 was activated by the alpha subunit of Gq but not by the transducin alpha subunit. However, unlike other PLC-beta isozymes, PLC-beta 4 was not responsive to activation by G beta gamma subunits.
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PMID:Regulation of phospholipase C-beta 4 by ribonucleotides and the alpha subunit of Gq. 792 27

We studied the relative ability to activate phospholipase C (PLC) of four Gs-coupled receptors expressed in L cells at different densities. Stable cell lines expressing various levels of the luteinizing hormone receptor (LHR), the type 2 vasopressin receptor (V2R), or the type 1 or type 2 beta-adrenergic receptor (beta 1- or beta 2AR) were isolated. The PLC activity was assessed by the measurement of free intracellular Ca2+ concentrations and the accumulation of inositol phosphates. We previously reported that, at 24,000 sites/cell, the LHR in L cells stimulated adenylyl cyclase by 10-fold over basal levels and PLC by 50% over basal levels. The EC50 for stimulation was 20-fold higher for PLC than for adenylyl cyclase. We now report that LHR tends to stimulate PLC more at a higher receptor density and less at a lower density. EC50 values for accumulation of inositol phosphates remained unchanged. The human V2R and the human beta ARs are strong adenylyl cyclase stimulators, and their potential for dual signaling was unknown. Expressing the V2R at 100,000 sites/cell or more and the beta ARs at 300,000 sites/cell resulted in stimulation of PLC by these receptors. As with the LHR, higher concentrations of vasopressin or isoproterenol were needed to reach 50% stimulation of PLC, compared with that of adenylyl cyclase. The beta 1AR was a stronger PLC stimulator than was the beta 2AR. The orders of potency for isoproterenol, epinephrine, and norepinephrine to stimulate adenylyl cyclase and PLC were the same for each of the two beta ARs. These results indicate that the ability of Gs-coupled receptors to stimulate PLC is dependent on the levels of receptor expression, and they suggest that dual signaling potential is a common property of Gs-coupled receptors and possibly also of G(i)-coupled receptors.
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PMID:Dual signaling potential is common among Gs-coupled receptors and dependent on receptor density. 793 26

1. Metabotropic glutamate receptor subtype 1 (mGluR1), when expressed in Xenopus oocytes, activates phosphoinositide-specific phospholipase C (PLC) in a G protein-dependent manner. This reaction results in the activation of chloride channels in the oocytes, and can be monitored electrophysiologically. We expressed different G protein alpha-subunits together with mGluR1 in oocytes, and examined the effects of these G protein subunits on the PLC-mediated reaction. 2. The expression of the alpha-subunit of GL2, a bovine version of G11, which is a member of the Gq subgroup, potentiated the mGluR1-evoked reaction, whereas the alpha-subunit of GL1, a bovine G14, which is also a member of the Gq subgroup, strongly suppressed it. The expression of Gs alpha also suppressed this reaction. 3. We then expressed G beta 1 gamma 2-subunits in addition to the G alpha-subunits, and examined the mGluR1-evoked reactions. Both the potentiation and suppression by GL2 alpha and GL1 alpha, respectively, were more pronounced in the presence of the G beta 1 gamma 2-subunits. In contrast, the suppression by Gs alpha was completely reversed by G beta 1 gamma 2. 4. The direct activation of G proteins by the intracellular injection of either fluoride ions or guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S) causes similar PLC-mediated reactions. The expression of GL2 alpha, GL1 alpha or Gs alpha caused potentiation, suppression and no change, respectively, on the fluoride- (or GTP gamma S-) evoked reactions.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:G protein-mediated inhibition of phosphoinositide metabolism evoked by metabotropic glutamate receptors in frog oocytes. 801 96

We evaluated the G protein selectivity of chimeric M1 and M2 muscarinic cholinergic receptors in which either the third intracellular (I3) loop or the N-terminal portion of this loop (the I3N peptide) was replaced by the corresponding sequence from the beta 1-adrenergic receptor. The chimeras retained agonist-dependent G protein regulatory activity, but were completely promiscuous among potential G protein targets. When expressed in transfected cells, the chimeric receptors activated adenylyl cyclase, the major target of the beta-adrenergic receptor, and activated phospholipase C via a pertussis toxin-insensitive G protein, presumably a Gq. Gs is not a target of either muscarinic receptor, and Gq is not a cellular target of either the M2 muscarinic or beta-adrenergic receptor. When co-reconstituted into phospholipid vesicles with purified G proteins, the chimeric receptors were completely nonselective among all G proteins tested. They activated Gi, G(o), Gz, and Gs with similar efficiencies. This promiscuity was largely suppressed, both in transfected cells and in reconstituted vesicles, by the additional replacement of the second intracellular (I2) loop of the beta-adrenergic receptor. Such double substitutions created receptors specific for Gs, the target of the beta-adrenergic receptor. These findings suggest that G protein specificity depends on the proper combination of multiple regions on a receptor's cytoplasmic surface. In addition, the promiscuous receptors described here may be useful for regulating novel G proteins whose natural regulators are not yet known.
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PMID:Chimeric muscarinic cholinergic:beta-adrenergic receptors that are functionally promiscuous among G proteins. 803 54

Phosphoinositide-specific phospholipase C (PI-PLC) catalyzes the hydrolysis of phosphatidylinositol 4,5-bisphosphate to inositol 1,4,5-trisphosphate (IP3) and diacylglycerol. IP3 induces the release of Ca2+ from intracellular stores, and diacylglycerol acts as the physiological activator of protein kinase C. Several distinct PI-PLC enzymes have been identified from various cells. Based on the primary sequences, PI-PLC isozymes are divided into three families: PLC-beta, PLC-gamma, and PLC-delta. Substantial evidence has strongly suggested that G proteins regulate PI-PLC in various cell-stimulation systems and that there might be two distinct pathways (pertussis toxin-sensitive and pertussis toxin-insensitive). Recently, it has become apparent that beta-type PLC isoforms are activated by the heterotrimeric G protein subfamily Gq. Careful studies using in vitro and in vivo reconstitution systems have further suggested that the alpha-subunits of Gq/11/16 specifically regulate PLC-beta 1 and PLC-beta 3 and that the beta gamma -subunits of the Gi subfamily interact with PLC-beta 2, which are considered to be responsible for the pertussis toxin-insensitive and the pertussis toxin-sensitive pathways, respectively. In this paper, involvement of G proteins in the regulation of phospholipase A2 and phosphatidylcholine-specific PLC and PLD is also discussed.
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PMID:[Phospholipid metabolism regulated by heterotrimeric G proteins]. 803 70

The small GTP-binding protein ARF has been shown recently to regulate phospholipase D (PLD). In order to investigate the role of ARF proteins in regulated exocytosis, we have used the N-terminal peptide ARF1(2-17) of the ARF1 protein. ARF1 reconstituted PLD activity in cytosol-depleted HL60 cells was inhibited by ARF1(2-17). In the presence of endogenous cytosol, ARF1(2-17) also inhibited GTP-gamma-S-stimulated PLD activity and exocytosis. Mastoparan Politses jadwagae and mastoparan Vespula lewisii which exhibit similar structural properties to ARF1(2-17) also inhibited GTP-gamma-S-stimulated PLD and exocytosis. GTP-gamma-S-stimulated phospholipase C-beta (PLC-beta) was also inhibited by ARF(2-17) and mastoparan. In cytosol-depleted HL60 cells, the ARF(2-17) inhibited the reconstitution of GTP-gamma-S-stimulated PLC-beta activity with exogenously-added PLC-beta 1 and phosphatidylinositol transfer protein. We conclude that the widely-used ARF1(2-17) peptide inhibits both ARF-independent (i.e. PLC-beta) and ARF-dependent pathways (i.e. PLD) and therefore cannot be regarded as a specific inhibitor of ARF function.
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PMID:ARF1(2-17) does not specifically interact with ARF1-dependent pathways. Inhibition by peptide of phospholipases C beta, D and exocytosis in HL60 cells. 804 98

The alpha T3-1 cell line, an immortalized gonadotroph cell line, expresses high levels of the gonadotrophin-releasing hormone (GnRH) receptor. Sustained exposure of these cells to the GnRH receptor agonist des-Gly10-[D-Ala6]luteinizing hormone-releasing hormone ethylamide resulted in a substantial down-regulation of cellular levels of a combination of the alpha subunits of the phospholipase C-beta 1-linked G proteins Gq and G11, as assessed by immunoblotting with an antiserum able to identify these two proteins equally. This effect was dependent upon the concentration of agonist used (EC50 = 4 nM) and on the time of the treatment (t1/2 = 6 hr) when a maximally effective concentration of agonist (1 microM) was used. Comparison of agonist regulation of inositol phosphate generation and Gq alpha/G11 alpha down-regulation demonstrated that effects on inositol phosphate production were approximately 3-fold more potent. In contrast to Gq alpha/G11 alpha, membrane-associated levels of Gs alpha and G12 alpha, the G proteins that transduce stimulatory and inhibitory regulation, respectively, of adenylyl cyclase, were not altered by agonist treatment. Analysis of mRNA by reverse transcriptase/polymerase chain reaction indicated the coexpression by alpha T3-1 cells of mRNA corresponding to both Gq alpha and G11 alpha. Immunoblotting with antisera selective for either Gq alpha or G11 alpha confirmed their coexpression. Resolution of membranes from untreated and agonist-treated alpha T3-1 cells under sodium dodecyl sulfate-polyacrylamide gel electrophoresis conditions able to separate Gq alpha from G11 alpha indicated that G11 alpha was more prevalent than Gq alpha at steady state but that agonist treatment regulated cellular levels of both of these G proteins in a nonselective manner. Sustained activation of protein kinase C with phorbol myristate acetate was unable to mimic agonist regulation of cellular Gq alpha/G11 alpha levels, as was treatment of alpha T3-1 cells with the selective protein kinase C inhibitor chelerythrine. These data suggest that the GnRH receptor is able to interact functionally with both Gq alpha and G11 alpha in alpha T3-1 cells and that sustained exposure to a GnRH receptor agonist selectively regulates the cellular levels of the G proteins that interact with the receptor.
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PMID:The gonadotrophin-releasing hormone receptor of alpha T3-1 pituitary cells regulates cellular levels of both of the phosphoinositidase C-linked G proteins, Gq alpha and G11 alpha, equally. 805 44


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