Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0043167 (pertussis)
 19,595 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Stimulation of leukocytes with chemoattractant ligands activates phospholipid turnover and calcium release, ultimately leading to chemotaxis, degranulation, and the inflammatory response. The leukocyte response to these ligands is transduced by the interaction of transmembrane receptors with GTP-binding regulatory proteins (G-proteins). To examine the mechanisms of signal transduction by these receptors, we transfected cDNA clones encoding the receptors for the active cleavage product of the fifth component of complement (C5a) and platelet-activating factor (PAF) into COS-7 cells, then measured the production of inositol phosphates (IP) in response to stimulation with these chemoattractant ligands. Cells transfected with the C5a receptor showed no increase in IP production when stimulated with ligand (5-120 nM). However, in cells co-transfected with these receptors and with the cDNA for G alpha 16, a G-protein alpha subunit that is specific to cells of hematopoietic lineage, addition of ligand caused up to a 5-fold increase in IP production. This interaction was specific, as co-transfection of receptors with the G-proteins G alpha q or G alpha 11 did not allow ligand-dependent increase in IP production. In contrast, ligand-dependent activation of IP production was seen in COS cells transfected solely with the PAF receptor. These results indicate that the C5a receptor utilizes signaling pathways distinct from the PAF receptor and suggest that a pertussis toxin-resistant G-protein, G alpha 16, may play a role in the leukocyte response to inflammatory ligands.
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PMID:Specific interactions of chemoattractant factor receptors with G-proteins. 848 84

At least 30 G protein-linked receptors stimulate phosphatidylinositol 4,5-bisphosphate phosphodiesterase (phospholipase C beta, PLC beta) through G protein subunits to release intracellular calcium from the endoplasmic reticulum (Clapham, D. E. (1995) Cell 80, 259-268). Although both G alpha and G beta gamma G protein subunits have been shown to activate purified PLC beta in vitro, G alpha q has been presumed to mediate the pertussis toxin-insensitive response in vivo. In this study, we show that G beta gamma plays a dominant role in muscarinic-mediated activation of PLC beta by employing the Xenopus oocyte expression system. Antisense nucleotides and antibodies to G alpha q/11 blocked the m3-mediated signal transduction by inhibiting interaction of the muscarinic receptor with the G protein. Agents that specifically bound free G beta gamma subunits (G alpha-GDP and a beta-adrenergic receptor kinase fragment) inhibited acetylcholine-induced signal transduction to PLC beta, and injection of G beta gamma subunits into oocytes directly induced release of intracellular Ca2+. We conclude that receptor coupling specificity of the G alpha q/G beta gamma heterotrimer is determined by G alpha q; G beta gamma is the predominant signaling molecule activating oocyte PLC beta.
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PMID:The G protein beta gamma subunit transduces the muscarinic receptor signal for Ca2+ release in Xenopus oocytes. 853 Apr 11

We have previously shown that the high-affinity binding of substance P (SP) to its receptor is dependent on an interaction with a PTX-insensitive G protein. This G protein couples SP receptor activation to stimulation of its effector, phospholipase C. In this study, we combined photoaffinity labeling, chemical cross-linking techniques, and immunological characterization using sequence-specific antibody probes to identify G proteins that couple to the SP receptor. First we covalently labeled the SP receptor present on rat submaxillary gland membranes with a radioiodinated photoreactive derivative of SP, p-benzoyl-L-phenylalanine(8)-substance P (125I-[Bpa8]SP). Photoincorporation of this SP derivative was susceptible to guanine nucleotide inhibition, indicating that the receptor was coupled to its G protein during labeling. We then used a chemical cross-linking agent to covalently link the photoaffinity labeled SP receptor and its associated G protein. Cross-linking generated a 96 kDa product, formation of which was prevented by the addition of a guanine nucleotide, but not an adenine nucleotide, following photolabeling, but prior to cross-linking. Furthermore, the 96 kDa cross-linked complex was absent in membranes which had been depleted of G proteins by treatment with alkaline buffer prior to addition of the cross-linking agent. Reductive cleavage of the cross-link in the isolated 96 kDa complex yields two products: the 53 kDa SP receptor and a 42 kDa protein identified by immunoblot analysis as either G alpha q or G alpha 11. Antisera against a common sequence within G alpha s, G alpha i, and G alpha o showed no immunoreactivity to the complex or its cleavage products. These results provide the first direct evidence of specific interaction between photoaffinity labeled SP receptor and the alpha subunits of Gq and G11, members of a family of G proteins known to be associated with pertussis toxin-insensitive phospholipase C activation.
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PMID:Chemical cross-linking of the substance P (NK-1) receptor to the alpha subunits of the G proteins Gq and G11. 860 28

Parafollicular (PF) cells secrete 5-hydroxytryptamine in response to increased extracellular Ca2+ ([Ca2+]e). This stimulus causes Cl- channels in PF secretory vesicles to open, leading to vesicle acidification. PF cells express a plasmalemmal heptahelical receptor (CaR) that binds Ca2+, Gd3+, and Ba2+. We now report that the CaR mediates vesicle acidification. Ca2+, Gd3+, and Ba2+ induced vesicle acidification, which was independent of channel-mediated Ca2+ entry. Agonist-induced vesicle acidification was blocked by pertussis toxin, inhibitors of phosphatidylinositol-phospholipase C, calmodulin, NO synthase, guanylyl cyclase, or protein kinase G. PF cells contained NO synthase immunoreactivity, and vesicles were acidified by NO donors and dibutyryl cGMP. [Ca2+]e, and Gd3+ mobilized thapsigargin-sensitive internal Ca2+ stores. [35S]G alpha i and [35S]G alpha q were immunoprecipitated from PF membranes incubated with agonists in the presence of [35S]adenosine 5'-O-(thiotriphosphate). Labeling of G alpha i but not G alpha q was antagonized by pertussis toxin. Vesicles acidified in response to activation of protein kinase C; however, protein kinase C inhibition blocked calcium channel- but not CaR-dependent acidification. We propose the following signal transduction pathway: CaR -> Gi -> phosphatidylinositol-phospholipase C -> inositol 1,4,5-trisphosphate -> [Ca2+]i -> Ca2+/calmodulin -> NO synthase -> NO -> guanylyl cyclase -> cGMP -> protein kinase G -> opens vesicular Cl- channel.
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PMID:Acidification of serotonin-containing secretory vesicles induced by a plasma membrane calcium receptor. 862 45

Guanine nucleotide binding proteins (G proteins) have been implicated in the pathophysiology of bipolar affective disorder. In the present investigation receptor-mediated G protein activation and changes in G protein trimeric state were examined in frontal cortical membranes obtained from postmortem brains of bipolar affective disorder subjects and from age-, sex-, and postmortem interval-matched controls. Stimulation of cortical membranes with serotonin, isoproterenol, or carbachol increased guanosine 5'-O-(3-[35S]thiophosphate) ([35S]GTP gamma S) binding to specific G alpha proteins in a receptor-selective manner. The abilities of these receptor agonists to stimulate the binding of [35S]GTP gamma S to the G alpha proteins was enhanced in membranes from bipolar brains. Immunoblot analyses showed increases in the levels of membrane 45- and 52-kDa G alpha S proteins but no changes in the amounts of G alpha i, G alpha o, G alpha Z, G alpha q/11, or G beta proteins in membrane or cytosol fractions of bipolar brain homogenates. Pertussis toxin (PTX)-activated ADP-ribosylations of G alpha i and G alpha o were enhanced by approximately 80% in membranes from bipolar compared with control brains, suggesting an increase in the levels of the trimeric state of these G proteins in bipolar disorder. Serotonin-induced, magnesium-dependent reduction in PTX-mediated ADP-ribosylation of G alpha i/G alpha o in cortical membranes from bipolar brains was greater than that observed in controls, providing further evidence for enhanced receptor-G protein coupling in bipolar brain membranes. In addition, the amounts of G beta proteins that coimmunoprecipitated with the G alpha proteins were also elevated in bipolar brains. The data show that in bipolar brain membrane there is enhanced receptor-G protein coupling and an increase in the trimeric state of the G proteins. These changes may contribute to produce exaggerated transmembrane signaling and to the alterations in affect that characterize bipolar affective disorder.
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PMID:Receptor-mediated activation of G proteins is increased in postmortem brains of bipolar affective disorder subjects. 875 21

The action of angiotensin II (ANG II) was studied in single myocytes from rat portal vein, in which the cytoplasmic Ca++ concentration was estimated by emission from fluorescent dyes and the Ca++ channel current was measured with the whole-cell mode of the patch-clamp technique. ANG II stimulated Ca++ channel current through L-type Ca++ channels and initiated a slow and small increase in the cytoplasmic Ca++ concentration in cells in which intracellular Ca++ stores had been depleted by pretreatment with ryanodine and caffeine. Both Ca++ channel current stimulation and Ca++ responses were selectively inhibited by losartan, indicating activation of angiotensin AT1 receptors. Activation of Ca++ channels by ANG II was insensitive to treatment with pertussis toxin and cholera toxin. Intracellular applications of anti-G alpha q/alpha 11 and anti-phosphatidylinositol antibodies had no effect on the ANG II-induced stimulation of Ca++ channel current, indicating that phosphatidylinositol-specific phospholipase C was not involved in this signaling pathway. Down-regulation of protein kinase C and application of an inhibitor of protein kinase C blocked the ANG II-induced effects. Tricyclodecan-9-yl xanthogenate (an inhibitor of non-phosphatidylinositol-specific phospholipases C and phospholipases D) but not propranolol (an inhibitor of phospholipase D-derived diacylglycerol formation) suppressed the ANG II-induced effects. These data suggest that phosphatidylcholine-specific phospholipase C is involved in the ANG II signaling pathway leading to stimulation of L-type Ca++ channels by protein kinase C.
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PMID:Angiotensin II-mediated activation of L-type calcium channels involves phosphatidylinositol hydrolysis-independent activation of protein kinase C in rat portal vein myocytes. 876 93

Using acutely isolated rat substantia nigra neurons, our previous studies indicated that sulfated cholecystokinin octapeptide (CCK-8) excites substantia nigra dopaminergic neurons by increasing the cationic conductance and that pertussis toxin-insensitive G proteins mediate CCK-8 induction of cationic currents. G alpha q and G alpha 11 are expressed in various tissues, including the brain, and likely to mediate pertussis toxin-insensitive neural signal transductions. In the present study, two different experiments were performed to test the hypothesis that G alpha q/11 mediates CCK-8 enhancement of the cationic conductance. First, we investigated the expression of G alpha q and G alpha 11 mRNAs in CCK-8-responsive substantia nigra dopaminergic neurons by combining whole-cell patch-clamp recordings with a single-cell reverse transcriptase-polymerase chain reaction assay. After CCK-8-evoked cationic currents were recorded, cellular RNA was harvested from single neurons and used as a template for the subsequent reverse transcriptase-polymerase chain reaction analysis. G alpha q and G alpha 11 mRNAs were present in all substantia nigra dopaminergic neurons that responded to CCK-8. Substantia nigra dopaminergic neurons were also internally perfused with the antibody raised against the common C-terminus of G alpha q and G alpha 11 during whole-cell recordings. CCK-8 failed to induce cationic currents after dopaminergic neurons were dialyzed with the anti-G alpha q/11 antibody. Our studies suggest that CCK-8 activation of the cationic conductance in substantia nigra dopaminergic neurons is transduced by G alpha q and/or G alpha 11.
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PMID:G alpha q/11 mediates cholecystokinin activation of the cationic conductance in rat substantia nigra dopaminergic neurons. 876 67

Bombesin stimulation of inositol 1,4,5-trisphosphate (Ins P3) formation in rat sonicated pancreatic acinar cells was inhibited by an antibody directed against the pertussis toxin (PTX)-sensitive GTP-binding G alpha i3 protein but not by an anti-G alpha q-11 antibody. After solubilization and gel filtration, [125I-Tyr4]bombesin binding sites were recovered in a peak of protein of 67 approximately 90 kDa with a maximal enrichment corresponding to a molecular mass of 83-kDa. Results obtained from the non-hydrolysable GTP analog guanosine-5'-[gamma-thio]triphosphate (GTP gamma S) binding, PTX-stimulated ADP-ribosylation and immunoblotting showed that the 83-kDa fraction contained the G alpha i3 protein but not the G alpha q-11 protein. Furthermore, GTP gamma S increased the bombesin binding dissociation constant (KD) from 0.32 to 0.60 nM, while the anti-G alpha i3 antibody decreased the maximal binding capacity (Bmax) from 50 to 25 fmol/mg protein without affecting the KD. Mixing solubilized bombesin binding sites with a phospholipase C (PLC) preparation from rat pancreas reconstituted a bombesin-stimulated PLC activity which was markedly inhibited by the anti-G alpha i3 antibody but unaffected by the anti-G alpha q-11 antibody. In addition, this stimulation was inhibited by an anti-PLC beta 1 antibody. This result supports the involvement of the PLC beta 1 isoform in bombesin receptor activation.
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PMID:Bombesin activation of phospholipase C beta 1 in rat acinar pancreatic cells involves the pertussis toxin-sensitive G alpha i3 protein. 879 79

1. We sought to reconstitute and characterize G-protein linked phosphatidyl-D-inositol 4,5-bisphosphate (PIP2)-directed phospholipase C (PLC) isoform activity in pig aortic vascular smooth muscle. 2. Six soluble PLC isoforms, namely gamma 1, delta 1 and beta 1 to beta 4 were partially separated by heparin affinity chromatography and were identified by Western blotting using specific antibodies. 3. In separate experiments, PLC activity was measured in the eluted fractions. Four of the partially resolved PLC isoforms gamma 1, beta 4, beta 2 and beta 1, showed corresponding activity using exogenous [3H]-PIP2 as substrate. 4. The isolated soluble PLC isoforms were reconstituted with receptors and guanyl nucleotide regulatory proteins (G-proteins) by addition of plasma membranes, the phospholipids which had been prelabelled with [3H]-myo-inositol. When so reconstituted PLC beta 2, beta 3 and beta 4 were inhibited (40 +/- 9, 47 +/- 12 and 40 +/- 5% respectively n = 12, +/-s.e.mean and each P < 0.05) by the addition of 1 mM guanosine 5'[beta gamma-imido]triphosphate (p[NH]ppG). 5. By contrast, when plasma membranes were preincubated with pertussis toxin to inhibit the activity of G-protein subunits G alpha i/alpha o the activities of PLC beta 2, beta 3 and beta 4 were stimulated (46 +/- 11, 31 +/- 9 and 37 +/- 8% respectively, n = 12, +/- s.e.mean and each P < 0.05) by the addition of p[NH]ppG. 6. Using well resolved fractions containing only PLC beta 3, time-dependent activity in the presence of p[NH]ppG was measurable only with membranes pretreated with pertussis toxin. 7. PLC beta 3 activity, measured with pertussis pretreated membranes, showed a dose-dependent increase in the presence of p[NH]ppG or guanosine 5'-[gamma-thio]triphosphate (GTP[S]). This increase with 10 microM p[NH]ppG or GTP[S] 10% +/- 4 and 12% +/- 5 respectively (both P < 0.05 vs control without GTP analogue +/- s.e.mean, n = 10) was abolished by 50 microM guanosine 5'-[beta-thio]diphosphate (GDP[S]) which also reduced constitutive PLC beta 3 activity by 9% +/- 4. 8. G-protein antibodies were used to neutralize PLC activity. Antibody to G alpha q/alpha 11, added to membrane fractions pretreated with pertussis toxin and assayed with GTP[S], reduced PLC beta 3 activity by 21% +/- 6 P < 0.02, n = 6, but was without effect on non-pertussis pretreated membranes. Antibodies to G alpha i1/alpha i2 had no effect. Antibodies to G-protein beta subunits had no effect on PLC beta 3 activity with pertussis pretreated preparations but activity without pertussis pretreatment was increased by 30% +/- 10, P < 0.03, n = 6. All results were expressed as % change from controls containing rabbit IgG. 9. In conclusion, pig aortic vascular smooth muscle contains six PLC isoforms. Activation of pertussis sensitive G-protein by GTP analogues results in inhibition of PLC beta 3 activity from liberated G-protein beta gamma subunits. Stimulation of PLC beta 3 activity is associated with a G-protein of the G alpha q family acting through the alpha subunit. The results suggest that the G-protein linked PLC beta isoforms in vascular smooth muscle demonstrate dual regulation by an inhibitory pertussis-sensitive pathway and a stimulatory G-protein of the G alpha q family, which is the case for PLC beta 3. This dual regulation is analogous to that of adenyl cyclase.
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PMID:Phospholipase C isoforms in vascular smooth muscle and their regulation by G-proteins. 879 75

The objective of our study was to investigate if there are abnormalities in signal transducing G proteins in patients with panic disorder. We utilized selective antibodies to quantitate the levels of the G protein alpha subunits that regulate adenylyl cyclase activity (G alpha s and G alpha i2) and phosphoinositide turnover (G alpha q/11) in platelet membranes (and leukocyte membranes for G alpha s), and also carried out pertussis toxin (PT) catalyzed [32P]ADP-ribosylation in platelet membranes from a group of 13 untreated panic disorder patients, 10 untreated social phobia patients, and 12 healthy subjects. There were no significant differences among the three groups in the immunolabeling of G alpha s in platelets or leukocytes, or in the immunolabeling of G alpha i1/2, G alpha q/11, or PT-catalyzed [32P]ADP-ribosylation in platelets. Within the constraints imposed by using peripheral blood cells to reflect brain composition, our results do not provide support for G protein abnormalities in patients with panic disorder. These results contrast with those obtained using identical methodology in bipolar affective disorder, where elevated G alpha s in leukocytes has been reported (Manji et al. 1995).
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PMID:G-protein level quantification in platelets and leukocytes from patients with panic disorder. 884 Mar 54


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