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

Thyroliberin (TRH), vasoactive intestinal peptide (VIP) and somatostatin (SRIF) act through receptors that are coupled to guanine nucleotide-binding regulatory proteins (G proteins). Regulation of hormone action may occur at the level of G protein coupling to the receptor or effector systems. In this study we demonstrate that prolonged exposure (for up to 48 hr) of cultured rat pituitary adenoma GH3 cells to these hormones caused homologous and to some extent heterologous attenuation of the adenylyl cyclase (AC) (EC 4.6.1.1) responsiveness. In addition, TRH and SRIF diminished both TRH- and guanosine 5'-[beta gamma-imido]-triphosphate-enhanced phospholipase C (PLC) (EC 3.1.4.3) activity within the same time-course. Measurements of cells membrane levels of Gs protein alpha-subunit (Gs alpha), G(i)-1 alpha/G(i)-2 alpha, G(i)-3 alpha, G(o) alpha and G beta by immunoblotting were performed. TRH and VIP upregulated levels of all G proteins except G(o) alpha and G beta. In contrast, SRIF caused a marked reduction of G beta levels. Thus, TRH and VIP, both acting through Gs, both modulated the alpha-subunit levels of this signal transducer, whereas SRIF, which possibly acts through G(i)-2, did not change the steady state level of G(i)-2 alpha. The actions of TRH, VIP and SRIF are multifaceted at the G protein level, where modulations of subtypes not directly involved in their actions may occur. These findings emphasize the complexity expected to be found in the in vivo situation.
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PMID:Hypothalamic hormones modulate G protein levels and second messenger responsiveness in GH3 rat pituitary tumour cells. 135 62

We have investigated the possibility that adenylyl cyclase (AC) activity and membrane protein levels of the alpha-subunits of the stimulatory and inhibitory G-proteins of AC (Gs alpha and G(i)-2 alpha) in cultured prolactin-producing rat pituitary adenoma cells (GH3 cells) are modulated by phospholipase C (PLC)-generated second messengers. Pretreatment of cells (6-48 h) with ionomycin (1 microM) or 1-oleoyl-2-acetylglycerol (OAG; 1 microM) showed that ionomycin regulated Gs alpha levels in a time-dependent, biphasic manner; a two-fold increase followed a 40% initial reduction, while OAG lowered Gs alpha levels by more than 50% at all time-points. G(i)-2 alpha levels remained unchanged by both pretreatments. OAG, but not ionomycin, increased basal AC activity without increasing enzyme protein levels. Alterations in AC responsiveness to peptide hormones (e.g. thyroliberin and vasoactive intestinal peptide) correlated to membrane Gs protein alpha-subunit content. These results demonstrate the involvement of G-protein translation regulation as one mechanism of 'cross-talk' between the PLC- and AC-dependent signalling pathways.
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PMID:'Cross-talk' between phospholipase C and adenylyl cyclase involves regulation of G-protein levels in GH3 rat pituitary cells. 148 64

The presence of the pertussis toxin (PTX) insensitive GTP-binding proteins (C-proteins) G(q) alpha and/or G(11) alpha has been demonstrated in three different prolactin (PRL) and growth hormone (GH) producing pituitary adenoma cell lines. Immunoblocking of their coupling to hormone receptors indicates that G(q) and/or G(11) confer throliberin (TRH) responsive phospholipase C (PL-C) activity in these cells. The contention was substantiated by immunoprecipitation analyses showing that anti G(q)/11 alpha-sera coprecipitated PL-C activity. In essence, only G(q)/11 (but neither G(12) G(13) nor G(o)) seems to mediate the TRH-sensitive PL-C activity, while G(o) may be coupled to a basal or constitutive PL-C activity. Immunoblocking studies imply that the B gamma-complex also, to some extent, may stimulate GH(3) pituitary cell line PL-C activity. Finally, the steady state levels of G(q)/(11) alpha mRNA and protein were down regulated upon long term exposure of the GH(3) cells to TRH (but not to vasoactive intestinal peptide = VIP).
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PMID:Phospholipase C activation in rat pituitary adenoma (GH) cells. 886 41

Ghrelin, a 28 amino acid, octanoylated peptide, is an endogenous ligand for the growth hormone secretagogue receptor (GHS-R). In addition to various endocrine functions, including stimulation of GH release, ghrelin has been characterized as an important regulator of energy homeostasis. Ghrelin administration has been shown to increase adiposity in rodents and stimulate food intake in humans. Studies suggest that these orexigenic effects are mediated primarily through GHS-R expression in hypothalamic and pituitary neuronal pathways. In this context, GHS-R has been recognized as a potential target for the treatment of GH deficiency and body weight disorders. Cell lines provide convenient in vitro systems to identify and characterize potential pharmacophores and to analyze GHS-R functional activity. While recombinant cell lines that overexpress GHS-R have served as effective research tools for these studies, such cell lines may differ in signaling response to ghrelin compared with hypothalamic or pituitary cells expressing GHS-R. We show here that a cell line derived from a rat anterior pituitary adenoma, RC-4B/C, expresses endogenous GHS-R as judged by reverse transcriptase-PCR. In a Ca(2+)mobilization assay, RC-4B/C cells demonstrate a dose-dependent increase in intracellular [Ca(2+)] on stimulation with rat ghrelin and a related peptide agonist, hexarelin (EC(50), 1.0 nM and 1.7 nM respectively), but are unresponsive to treatment with inactive des-octanoyl rat ghrelin. A subclone, RC-4B/C.40, with a more robust and stable ghrelin response, was isolated from the parental population of cells to allow further analysis of GHS-R signal transduction. Using pertussis toxin and the phospholipase C inhibitor U-73122, we show that ghrelin signals through the Gq pathway in the RC-4B/C.40 cells. We also demonstrate that the ghrelin-induced rise of intracellular [Ca(2+)] in RC-4B/C.40 cells involves initial Ca(2+)release from intracellular stores followed by a sustained elevation that occurs via influx of extracellular Ca(2+) through ion channels. In addition, unlike observations reported in recombinant cell systems, the RC-4B/C.40 cells do not exhibit a high level of GHS-R constitutive activity as determined in a phosphatidylinositol hydrolysis assay. Overall, the data presented here suggest that the RC-4B/C parental and RC-4B/C.40 cells provide novel in vitro systems for the characterization of GHS-R pharmacophores and ghrelin signaling.
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PMID:Characterization of ghrelin receptor activity in a rat pituitary cell line RC-4B/C. 1690 23