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)

Mastoparan has been reported to induce a wide variety of cellular actions by activating GTP-binding proteins (G proteins) in various cells. Here, we demonstrate that mastoparan is able to stimulate the secretion of PRL from rat anterior pituitary tumor GH3 cells in dose- and time-dependent manners. Mastoparan had no effect on the accumulation of intracellular cAMP; however, it induced a rapid increase in the intracellular Ca2+ concentration in GH3 cells. Extracellular Ca2+ was required for mastoparan-induced PRL secretion, which was inhibited by nifedipine, an L-type Ca2+ channel blocker. Incubation of mastoparan with myo-[3H]inositol-labeled GH3 cells also resulted in the increased formation of inositol phosphates (InsPs) compared with control cells. Neomycin sulfate and U73122, both phospholipase C inhibitors, suppressed mastoparan-induced PRL secretion. Guanosine 5'-1beta-thioldiphosphate (GDPbetaS) encapsulated in GH3 cells by reversible electropermeabilization suppressed the response to mastoparan. However, pretreatment with pertussis toxin had no effect on the stimulation of PRL secretion by mastoparan, and both Mas7 (a highly active analogue of mastoparan) and Mas17 (an inactive analogue) enhanced the secretion of PRL to a similar level to that of mastoparan-induced GH3 cells. In contrast, the substance P-related peptide GPant-2A, a Gq antagonist, inhibited mastoparan-induced PRL release, whereas GPant-2, a G(i/o) antagonist, did not in electropermeabilized GH3 cells. Moreover, a specific G(q/11) antibody against the carboxyl terminus of the G(q/11) alpha-subunit blocked the stimulatory effect of mastoparan on secretion and mastoparan-stimulated InsPs production in digitonin-permeabilized GH3 cells. These results indicate that mastoparan induces the Ca2+-regulated secretion of PRL from GH3 cells by activating G(q/11) and the phospholipase C pathway.
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PMID:Mastoparan-stimulated prolactin secretion in rat pituitary GH3 cells involves activation of Gq/11 proteins. 911 92

1. The signal transduction mechanism mediating extracellular adenosine 5'-triphosphate (ATP)-induced calcium release in a renal epithelial cell line (A6) was investigated using the whole-cell voltage-clamp technique and fura-2 fluorescence measurement. 2. ATP (10 microM) activated calcium-dependent non-selective cation channels in cells held under voltage clamp. 3. Guanosine 5'-O-(2-thiodiphosphate) (GDP beta S; 0.1-1.0 mM) in the pipette inhibited the ATP-activated calcium-dependent currents. With guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S; 0.1-1.0 mM) in the pipette, the currents were spontaneously elicited without application of ATP. Pretreatment with pertussis toxin (PTX) affected neither the ATP-activated currents nor the increase in intracellular free calcium concentration ([Ca2+]i) evoked by ATP. 4. Intracellular application of neomycin or heparin inhibited the ATP-activated currents. Inositol 1,4,5-trisphosphate (IP3; 0.1-100 microM) in the internal solution produced currents similar to those due to ATP activation. 5. These results suggest that a PTX-insensitive guanosine 5'-triphosphate (GTP)-binding regulatory protein (G. protein) is involved in extracellular. ATP-induced phosphoinositide turnover and subsequent calcium release from IP3-sensitive stores, which subsequently activates the calcium-dependent channels in A6 cells.
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PMID:Calcium release from intracellular stores evoked by extracellular ATP in a Xenopus renal epithelial cell line. 926 16

Despite a great deal of research, the second messenger coupling of the dopamine D3 receptor has not yet been clearly established. The closely related D2 and D4 receptors have been shown to inhibit adenylyl cyclase activity in a variety of cell types, but the D3 receptor has little or no effect on this second messenger system. We now demonstrate that when the D3 receptor and adenylyl cyclase type V are coexpressed in 293 cells, the agonist quinpirole causes 70% inhibition of forskolin-stimulated cAMP levels. This effect seems to be selective for this adenylyl cyclase isoform because the D3 receptor does not inhibit adenylyl cyclase types I or VI and only weakly stimulates adenylyl cyclase type II. In contrast, the D2 receptor inhibits cAMP accumulation in 293 cells in the absence of cotransfected adenylyl cyclases and stimulates adenylyl cyclase type II to a greater extent than the D3 receptor. The inhibition of adenylyl cyclase type V by the D3 receptor is sensitive to pertussis toxin, suggesting the involvement of G proteins of the Gi family. Guanosine-5'-O-(3-thio)triphosphate binding studies indicate that the D3 receptor weakly activates all three Gialpha subunits, whereas the D2 receptor activates these G proteins to a substantially greater extent. However, despite its relative inability to promote G protein activation, the D3 receptor is capable of substantial and consistent inhibition of adenylyl cyclase type V. The robust second messenger coupling of the D3 receptor in a heterologous system with defined components provides a system for further studies of the function of this receptor and should facilitate the development and characterization of new D3 receptor ligands.
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PMID:Selective inhibition of adenylyl cyclase type V by the dopamine D3 receptor. 928 14

[35S]Guanosine-5'-O-(3-thio)triphosphate ([35S]GTPgammaS) binding to G proteins was measured by in vitro autoradiography in guinea pig and rat brain sections after activation by 5-hydroxytryptamine (5-HT) receptor agonists. 5-Carboxamidotryptamine stimulated binding strongly in hippocampus and lateral septum and weakly in substantia nigra. This effect was blocked in the substantia nigra by the 5-HT1B/1D receptor antagonist GR-127,935 and in the former two regions by the 5-HT1A antagonist NAN-190. 5-HT1B/1D receptor agonists stimulated binding in substantia nigra and in areas containing 5-HT1A receptors. In guinea pig substantia nigra, 5-(nonyloxy)-tryptamine maximally stimulated [35S]GTPgammaS binding by 54%, with an EC50 value of 62 nM; at 100 microM, this agonist increased binding by approximately 200% in hippocampus (with a 2-fold weaker EC50 value). The distribution of [3H]8-OH-DPAT binding sites was identical to that of the [35S]GTPgammaS labeling stimulated by the 5-HT1A agonist (R)-8-hydroxy-2-dipropylaminotetralin [(R)-8-OH-DPAT)]. (R)-8-OH-DPAT, (S)-8-OH-DPAT, and buspirone stimulated [35S]GTPgammaS binding in hippocampus by 340%, 140%, and 78%, with EC50 values of 71, 51, and 132 nM. Enhanced [35S]GTPgammaS binding was not detected in the presence of 5-HT1F, 5-HT2, 5-HT4, and 5-HT7 receptor agonists. Because activation of mu-opioid, muscarinic M2, histamine H3, and cannabinoid receptors was also visualized successfully, these data suggest that only receptors coupled to pertussis toxin-sensitive G proteins can be seen by [35S]GTPgammaS binding autoradiography. This study also shows that different 5-HT receptors coupled to these proteins can show a wide range of [35S]GTPgammaS binding stimulation. Although the functional significance of these variations is unclear, this technique offers advantages over receptor autoradiography because it does not require high affinity radioligands and provides a measure of agonist efficacies in various brain regions.
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PMID:5-Hydroxytryptamine1A and 5-hydroxytryptamine1B receptors stimulate [35S]guanosine-5'-O-(3-thio)triphosphate binding to rodent brain sections as visualized by in vitro autoradiography. 938 25

An immunization program against diphtheria has been implemented in Taiwan since 1955, using combined diphtheria, pertussis and tetanus (DPT) vaccine. Diphtheria immunoglobulin (DIG) level was assessed in serum samples obtained from 1138 children, aged 3-6 years from north, south, east and central part of Taiwan by the VERO cell neutralization method. Specimens were collected by simple random sampling of residents from Hsinchu, Taichung, Pingtung and Hwalien counties, including both aborigines and non-aborigines. The former lived in one or two villages in each county, and the latter lived in a single village next to the former. Ninety-five percent (1086/1138) had a DIG titre > or = 0.01 IU/ml. There was no significant difference by sex, or by residential area. Seventy-nine percent (901/1138) of the children had completed the primary immunization schedule (at the age of 2, 4, 6 and 18 months), and the prevalence of DIG titre > or = 0.1 IU/ml considered to be long-term protective was as follows: 74.6% for 3-year group; 74.5% for 4-year group; 67.9% for 5-year group; 84.7% for 6-year group (including 52.2% who had had a booster shot at early primary school). These findings show that the diphtheria vaccination program provides good immunity in childhood.
Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi 1995 Feb
PMID:Immunity to diphtheria in children three-to-six year of age in four counties of Taiwan. 977 82

Muscarinic receptor regulation of guanine nucleotide turnover on Gi/Go proteins in ventricular sarcolemma was investigated. In the absence of a muscarinic receptor (MR) agonist, GTP bound to background sites with a Kapp value of 60 nM and a Bmax of 50 pmol/mg. The addition of the MR agonist, carbachol, further increased GTP binding by 50 pmol/mg to sites with the same Kapp value of 60 nM. Pertussis toxin treatment reduced GTP binding to carbachol-regulated and background binding sites, thus identifying both sites as Gi/Go. The identity of the carbachol-regulated GTP binding sites was further confirmed by demonstrating that carbachol stimulated GTP binding and inhibited adenylyl cyclase with an EC50 value of 200 nM. Background and carbachol-regulated guanine nucleotide binding sites bound GDP with a Kapp value of 150 nM. However, maximal background GDP binding was 50 pmol/mg, whereas maximal carbachol-regulated GDP binding was only 12-15 pmol/mg. In sarcolemma preloaded with [3H]GDP, carbachol-regulated [3H]GDP release was strictly dependent on the presence of guanine nucleotides. The Kapp values for GTP and GDP to support carbachol-regulated [3H]GDP release were 60 nM and 150 nM, respectively. Guanosine 5'-O-(3-thiotriphosphate) (GDPbetaS) facilitated carbachol-regulated [3H]GDP release with a Kapp value of 2 microM. However, GTP was two times more efficacious than GDP or GDPbetaS in facilitating carbachol-regulated [3H]GDP release. Mn2+ also stimulated [3H]GDP release from carbachol-regulated sites by a mechanism not requiring guanine nucleotides. These studies indicate that two pools of muscarinic receptors, carbachol regulated and spontaneously active, regulate guanine nucleotide turnover on pertussis toxin sensitive Gi/Go. These studies further suggest that guanine nucleotide binding provides the signal to stimulate GDP release from receptor activated Gi/Go proteins. A quaternary mechanism involving G-protein interactions may be necessary to promote guanine nucleotide exchange on Gi/Go.
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PMID:Regulation of guanine nucleotide turnover on Gi/Go by agonist-stimulated and spontaneously active muscarinic receptors in cardiac membranes. 988 28

The effect of guanine nucleotide-binding protein (G protein) modifiers on the binding of the adenosine A2A receptor agonist 2-[4-(2-p-carboxyethyl[3H])phenyl-amino]-5'-N-ethylcarboxamidoadenosine ([3H]CGS 21680) and of the adenosine A1 receptor agonist [3H]R-phenylisopropyladenosine ([3H]R-PIA) to rat cortical and striatal membranes was studied. Guanosine 5'-(beta,gamma-imido)triphosphate (1-300 microM), which uncouples all G proteins, more effectively inhibited [3H]CGS 21680 (30 nM) binding in the cortex than [3H]R-PIA (2 nM) binding to cortical or striatal membranes or [3H]CGS 21680 (30 nM) binding in the striatum. N-Ethylmaleimide (1-300 microM) or pertussis toxin (1-100 microg/ml), which uncouple G(i)/G(o) protein-coupled receptors, more effectively inhibited [3H]R-PIA binding to cortical or striatal membranes and [3H]CGS 21680 binding in the cortex than [3H]CGS 21680 binding in the striatum. Cholera toxin (2.5-250 microg/ml), which uncouples G(S) protein-coupled receptors, more effectively inhibited [3H]CGS 21680 binding in the striatum than [3H]CGS 21680 binding in the cortex and less effectively inhibited [3H]R-PIA binding to cortical or striatal membranes. Treatment of solubilised cortical membranes with pertussis toxin (50 microg/ml) decreased [3H]CGS 21680 (30-100 nM) binding which almost fully recovered after reconstitution with G(i)/G(o) proteins. The K(i) for displacement of [2-3H]-(4{2-[7-amino-2-(2-furyl)(1,2,4)triazolo(2,3-a)(1,3,5)triazin+ ++-5-ylamino]ethyl}phenol) ([3H]ZM 241385, 1nM) by CGS 21680 was 110 nM (95%CI: 98-122 nM) in non-treated, 230 (167-292) nM in pertussis toxin (25 microg/ml)-treated and 222 (150-295) nM in cholera toxin (50 microg/ml)-treated cortical membranes; in contrast, the K(i) for displacement of [3H]-5-amino-7-(2-phenylethyl)-2-(2-furyl)-pyrazolo(4,3-e)-1,2,4-triazol o(1,5-c)pyrimidine ([3H]SCH 58261, 1 nM) by CGS 21680 was 74 (57-91) nM in non-treated, 71 (44-100) nM in pertussis toxin-treated and 147 (100-193) nM in cholera toxin-treated cortical membranes. Finally, CGS 21680 displaced monophasically the binding of the A1 antagonist, [3H]8-cyclopentyl-1,3-dipropylxanthine (2 nM), and the A1 agonist, [3H]R-PIA (2 nM), in 2 or 10 mM Mg(2+)-medium, either at 25 degrees C or 37 degrees C, in cortical or striatal membranes. These results indicate that CGS 21680 does not bind to A1 receptors and that limbic CGS 21680 binding sites differ from striatal-like A2A receptors since they couple to G(i)/G(o) proteins, as well as to G(s) proteins.
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PMID:G protein coupling of CGS 21680 binding sites in the rat hippocampus and cortex is different from that of adenosine A1 and striatal A2A receptors. 1034 28

Pertussis toxin (PT) is the major protective antigen of acellular pertussis vaccine (aP). We have established an optimal culture condition for the growth of B. pertussis and the production of PT in a laboratory scale fermentor. It was found that when the dissolved oxygen in medium was supplied with pure oxygen instead of air, the yield of PT was dramatically increased (i.e. from 2-3 mg/l using air to 8-10 mg/l using pure oxygen). PT was purified by affinity chromatography using hydroxyapatite and fetuin-sepharose columns. SDS-PAGE analysis and CHO cell clustering test showed that the purified PT was comparable to the reference PT in purity and biological activity. The purified PT could be detoxified by formaldehyde (d-PT). The results of CHO cell clustering neutralization assay and ELISA showed that the antibody induced by d-PT in mice was comparable to that induced by PT contained in a commercial DTaP. These results indicated that the immunogenicity of our d-PT was retained after the purification and detoxification procedures.
Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi 1997 May
PMID:Production and purification of Bordetella pertussis toxin. 1059 13

A study was undertaken to evaluate the safety and immunongenicity of a conjugate Haemophilus influenzae type b (Hib) vaccine (HibTITER) when administered concurrently with DTP (diphteria, tetanus and pertussis) vaccine in separate syringes. A total of 90 healthy children (45 per group) were randomized to receive either TETRAMUNE, a vaccine combining HibTITER with whole-cell DTP (group A), or DTP and HibTITER administered concurrently (group B) in separate syringes at approximately 2, 4 and 6 months of age in Taiwan. All children in group B achieved anti-Hib PRP (polyribosylribitol phosphate) antibody titers above 0.15 microgram/ml and 91% developed antibody titers above 1.0 microgram/ml following the third immunization. Incidences of adverse reactions were comparable between groups A and B. Besides, the incidences of adverse reactions were not significantly more frequent compared with DTP vaccination alone. We concluded that HibTITER was highly immunogenic and safe when administered concurrently with DTP vaccine to Taiwanese children. TETRAMUNE was also safe and the number of injections may be reduced in the future.
Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi 1997 May
PMID:Immunogenicity of Haemophilus influenzae b conjugate vaccine (HibTITER) and safety of HibTITER and a combination vaccine of diphtheria, tetanus, pertussis and HibTITER in infants two months of age: a preliminary report. 1059 15

Pertussis toxin (PT), a typical A-B oligomer exotoxin of Bordetella pertussis, has been demonstrated to be an essential protective antigen for acellular pertussis vaccine against whooping cough. In order to investigate the associated functionality ascribed to its components, we have purified A and B oligomers for the activity study. The A oligomer (S1 subunit) of PT was expressed in E. coli B834 (DE3) harboring expression vector (pET-20b) with the insert of S1 coding region and purified by metal-chelating column. The B oligomer was isolated by a single-step purification procedure. Individually, recombinant S1 and B oligomer exhibited quite distinct biological activities in vivo. S1 subunit induced leukocytosis-promoting (LP) activity, but did not affect mouse body weight-gain. On the contrary, B oligomer reduced mouse body weight-gain but did not reveal LP activity. In vitro, the combination of S1 subunit and B oligomer could enhance the toxic activities as exhibited by native PT and showed an additive toxicity in CHO cell clustering test and hemagglutination assay.
Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi 1997 Aug
PMID:Preparation and characterization of Pertussis toxin subunits. 1059 23


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