Journal of receptor research最新文献

Characterization of mammalian Gs-alpha proteins expressed in yeast. 哺乳动物gs - α蛋白在酵母中的表达。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101510

J M Stadel, D J Ecker, D A Powers, J Marsh, K Hoyle, M Gross, M D Minnich, T R Butt, S T Crooke

{"title":"Characterization of mammalian Gs-alpha proteins expressed in yeast.","authors":"J M Stadel, D J Ecker, D A Powers, J Marsh, K Hoyle, M Gross, M D Minnich, T R Butt, S T Crooke","doi":"10.3109/10799899409101510","DOIUrl":"https://doi.org/10.3109/10799899409101510","url":null,"abstract":"The guanine nucleotide regulatory protein, GS, mediates transmembrane signaling by coupling membrane receptors to the stimulation of adenylyl cyclase activity. The full length coding sequences for the M(r) = 42-45,000, short form (S), and M(r) = 46-52,000, long form (L), of the alpha-subunits of rat GS were placed in yeast expression vectors under the regulatory control of the copper-inducible CUP1 promoter and transformed into Saccharomyces cerevisiae. In the presence of 100 microM CuSO4, the transformed yeast expressed GS-alpha mRNAs and proteins. In reconstitution experiments, rat GS-alpha(S and L), solubilized from yeast membranes with 1% cholate, conferred NaF-, (-)isoproterenol-, and guanine nucleotide-dependent sensitivity to adenylyl cyclase catalytic units in S49 lymphoma cyc- cell membranes, which are devoid of endogenous GS-alpha. GS-alpha (S) demonstrated twice the activity of GS-alpha(L) in reconstitution assays of fluoride-stimulated adenylyl cyclase activity. Comparison of GS-alpha (S) expressed in yeast with GS purified from rabbit liver or human erythrocytes showed that the crude recombinant protein was fully competent in reconstituting NaF-stimulated adenylyl cyclase activity, but was only 2-5% as potent as purified GS. Addition of bovine brain beta gamma subunits during reconstitution enhanced all parameters of adenylyl cyclase activity for GS-alpha(S and L) obtained from yeast. In contrast, transducin beta gamma only enhanced agonist-stimulated adenylyl cyclase activity for GS-alpha (S and L) following reconstitution. These results demonstrate that the expression of functional mammalian GS-alpha subunits in yeast may be useful for their biochemical characterization.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 6-8","pages":"357-79"},"PeriodicalIF":0.0,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409101510","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18876859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Sarafotoxin-induced calcium mobilization in cultured dog tracheal smooth muscle cells. 萨拉福毒素诱导犬气管平滑肌细胞的钙动员。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101513

C M Yang, R Ong, J T Hsieh, Y L Yo

{"title":"Sarafotoxin-induced calcium mobilization in cultured dog tracheal smooth muscle cells.","authors":"C M Yang, R Ong, J T Hsieh, Y L Yo","doi":"10.3109/10799899409101513","DOIUrl":"https://doi.org/10.3109/10799899409101513","url":null,"abstract":"Sarafotoxin b (S6b)-induced changes in intracellular Ca2+ concentration ([Ca2+]i) were monitored in cultured canine tracheal smooth muscle cells (TSMCs) by a fluorescent Ca2+ indicator fura-2. S6b elicited an initial transient peak followed by a sustained elevation of [Ca2+]i. BQ-123, an endothelin-A (ETA) receptor antagonist, had a high affinity to block the rise in [Ca2+]i response to S6b. In the absence of external Ca2+, only an initial transient peak of [Ca2+]i was seen, the sustained elevation of [Ca2+]i could then be evoked by addition of 1.8 mM Ca2+. Ca2+ influx was required for the changes of [Ca2+]i, since the Ca(2+)-channel blockers, diltiazem, verapamil, and Ni2+, decreased both the initial and sustained elevation of [Ca2+]i in response to S6b. TSMCs pretreated with phorbol 12-myristate 13-acetate (PMA, 1 microM) for 30 min attenuated Ca2+ mobilization induced by S6b, which was reversed by staurosporine, a protein kinase C (PKC) inhibitor. The change of [Ca2+]i induced by S6b was attenuated by cholera toxin pretreatment, but not by pertussis toxin. These data demonstrate that the initial detectable increase in [Ca2+]i stimulated by S6b is due to the activation of ETA receptors and subsequent release of Ca2+ from internal stores, whereas the contribution of external Ca2+ follows and partially involves a diltiazem- and verapamil-sensitive process. The inhibition of PMA on S6b-induced Ca2+ mobilization was inversely correlated with membraneous PKC activity.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 6-8","pages":"423-45"},"PeriodicalIF":0.0,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409101513","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18876862","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

Expression of alpha-bungarotoxin receptor subtypes in chick central nervous system during development. 鸡中枢神经系统发育过程中α -班加罗毒素受体亚型的表达。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101508

C Gotti, M Moretti, R Longhi, L Briscini, B Balestra, F Clementi

引用次数: 24

In situ characterization of renal insulin receptors in the rat. 大鼠肾脏胰岛素受体的原位表征。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101509

L A Sechi, S De Carli, E Bartoli

{"title":"In situ characterization of renal insulin receptors in the rat.","authors":"L A Sechi, S De Carli, E Bartoli","doi":"10.3109/10799899409101509","DOIUrl":"https://doi.org/10.3109/10799899409101509","url":null,"abstract":"Insulin regulates carbohydrate metabolism, and water, sodium, potassium, and phosphate reabsorption in the kidney by binding to specific receptors. Insulin receptors have been identified in the kidney using membrane preparations obtained from both glomeruli and tubules. In this study, an autoradiographic technique was used to characterize insulin receptors in the rat kidney. Frozen tissue sections were preincubated to remove endogenously bound insulin, incubated in a buffer containing 200 microM 125I-Tyr-insulin, washed, and dried before exposure on Ultrofilm. Binding density was assessed by computerized microdensitometry. In the cortex, binding density was comparable in glomeruli and tubules. In the medulla, bound radioligand was found primarily in longitudinal structures traversing the outer portion, presumably corresponding to vascular bundles, and in the inner portion. Scatchard analysis of competition binding data resulted in curvilinear profiles, indicating either two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed one receptor site with Kd of 0.39 +/- 0.14 nmol/l and Bmax of 3.5 +/- 1.0 x 10(10) receptors/mm3 and another site with Kd of 0.30 +/- 1.1 pmol/l and a Bmax of 3.2 x 10(13) receptors/mm3. Thus, in situ autoradiography can be used to determine distribution and binding characteristics of insulin receptors in rat kidney and might be employed in receptor studies on rat models of human disease.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 6-8","pages":"347-56"},"PeriodicalIF":0.0,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409101509","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18876858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 27

Antibody binding to the juxtamembrane region of the insulin receptor alters receptor affinity. 抗体结合到胰岛素受体的近膜区域改变受体亲和力。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101511

D W Goodman, G Romero, P Isakson

{"title":"Antibody binding to the juxtamembrane region of the insulin receptor alters receptor affinity.","authors":"D W Goodman, G Romero, P Isakson","doi":"10.3109/10799899409101511","DOIUrl":"https://doi.org/10.3109/10799899409101511","url":null,"abstract":"The effect of three antibodies that interact with distinct regions of the insulin receptor (the alpha subunit (83-7), the juxtamembrane region near tyrosine 960 (960) or the carboxy terminal region of the beta subunit (CT-1) on insulin binding was examined. Detergent-solubilized insulin receptors from IM-9 cells immobilized on Sepharose beads by 960 antisera bound 2-3 times more 125I-insulin tracer (25-60 pM) than receptors immobilized with either 83-7 or CT-1. Pre-incubation of solubilized receptors with either 83-7 or 960 resulted in equivalent depletion (90%) of insulin binding activity from solubilized IM-9 cell extracts, suggesting that both antibodies were in excess and capable of binding a similar population of receptors. Antibody 960, but not CT-1 or 83-7, also increased insulin binding 2 fold to solubilized receptors precipitated with polyethylene glycol. To determine whether the altered binding observed with antibody 960 was due to increased affinity of the receptor for insulin or appearance of more insulin binding sites, binding studies were performed over a wide range of insulin concentrations. Analysis of the resulting binding curves indicated that 960 increased the affinity of the receptor for insulin 3 fold over control (kd = 0.3 nM for 960, and 0.9 nM for 83-7, respectively). The antibody 960 also specifically increased insulin binding to intact, saponin-permeabilized IM-9 cell membranes. These results indicate that binding of 960 antibody to the juxtamembrane region of the insulin receptor alters the affinity of the receptor for insulin. Since tyrosine 960 in the juxtamembrane region has been suggested to play a role in receptor signalling, changes in receptor conformation in this region that are likely to account for the change in affinity may play a role in signal transduction.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 6-8","pages":"381-98"},"PeriodicalIF":0.0,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409101511","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18876860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Isolation and characterization of neurokinin A receptor cDNAs from guinea-pig lung and rabbit pulmonary artery. 豚鼠肺和兔肺动脉神经激肽A受体cdna的分离与鉴定。

Journal of receptor research Pub Date : 1994-12-01 DOI: 10.3109/10799899409101512

D Aharony, J Little, C Thomas, S Powell, M Downey-Jones, A Graham

{"title":"Isolation and characterization of neurokinin A receptor cDNAs from guinea-pig lung and rabbit pulmonary artery.","authors":"D Aharony, J Little, C Thomas, S Powell, M Downey-Jones, A Graham","doi":"10.3109/10799899409101512","DOIUrl":"https://doi.org/10.3109/10799899409101512","url":null,"abstract":"cDNA clones for NK-2 receptors (NK-2R) were isolated from guinea-pig lung (GPl) and rabbit pulmonary artery (Rpa) using a polymerase chain reaction based methodology. The GPl NK-2R consists of 402 amino acids and encodes a protein with a relative molecular mass of 45,097. The Rpa NK-2R consists of 384 amino acids and encodes a protein with a relative molecular mass of 43,169. The GPl and Rpa NK-2Rs share significant amino acid sequence homology amongst themselves (90.1%), as well as with human, bovine, hamster and rat NK-2 receptors. The two receptors were stably transfected into mouse erythroleukemia cells, high-speed membranes were prepared from induced cells and their pharmacological properties examined utilizing [3H]-NKA in a receptor-binding assay. [3H]NKA bound to both NK-2Rs with high affinity (KD = 2-7 nM) and saturable (Bmax = 633-9000 fmol/mg protein) manner which was inhibited by GTP analogs. Competition experiments with agonists demonstrated identical order of potency in both NK-2Rs; NKA > [Nle10]NKA(4-10) > [beta-Ala8]NKA(4-10) > > Substance P > > > Senktide. Similarly, an identical profile for both receptors was observed with selective NK-2 antagonists: SR48,968 > MEN10,376 > > R396. The rank order of antagonist affinity is consistent with that in cloned human NK-2R and the observations of NK-2 receptor pharmacology in native human, guinea pig and rabbit tissues.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 6-8","pages":"399-421"},"PeriodicalIF":0.0,"publicationDate":"1994-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409101512","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18876861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Neurotensin binding to human embryonic lung fibroblasts. 神经紧张素与人胚胎肺成纤维细胞的结合。

Journal of receptor research Pub Date : 1994-07-01 DOI: 10.3109/10799899409066039

A Gupta, K A Hruska, K J Martin

{"title":"Neurotensin binding to human embryonic lung fibroblasts.","authors":"A Gupta, K A Hruska, K J Martin","doi":"10.3109/10799899409066039","DOIUrl":"https://doi.org/10.3109/10799899409066039","url":null,"abstract":"Several neuropeptides have been shown to regulate the function of cells involved in immune response and inflammation. Neurotensin is a 13 amino acid neuropeptide localized primarily to the nervous system and gut. Neurotensin also stimulates mast cell degranulation and enhances phagocytic and cytolytic capability of macrophages, suggesting that this peptide regulates inflammatory and immune responses. Fibroblasts play an important role in inflammation and tissue healing, and these processes may be regulated by several neuropeptides that have been shown to bind to fibroblasts. However neurotensin receptors have not been identified on fibroblasts. Human embryonic lung fibroblasts (HELF) were examined for binding and biological effects of neurotensin. 125I-neurotensin binding to adherent and confluent human embryonic lung fibroblasts (HELF), plated in 12mm diameter wells was specific and saturable. Computer-assisted resolution of the binding data demonstrated two classes of binding sites: a high affinity, low capacity site (Kd = 1.6 x 10(-11) M, 19.5 x 10(7) sites/well), and a low- affinity, high-capacity site (Kd = 10(-8) M, 4 x 10(9) sites/well). Neurotensin stimulated immediate, transient, dose-dependent increases of cytosolic calcium in HELF (threshold dose: 10(-11) M), suggesting release of calcium from intracellular stores. The novel finding of neurotensin receptors on fibroblasts provides further support for this neuropeptide's role as a regulator of inflammatory and immune responses.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 5","pages":"307-17"},"PeriodicalIF":0.0,"publicationDate":"1994-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409066039","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18817635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Dexamethasone mediated stabilization of insulin receptor mRNA. 地塞米松介导的胰岛素受体mRNA稳定。

Journal of receptor research Pub Date : 1994-07-01 DOI: 10.3109/10799899409066038

D Hines, V Hug, J R Levy

引用次数: 11

Surface distribution of the EGF receptor during differentiation of the human colon carcinoma cell line HT29-D4. 人结肠癌细胞系HT29-D4分化过程中EGF受体的表面分布。

Journal of receptor research Pub Date : 1994-07-01 DOI: 10.3109/10799899409066040

M Lehmann, M Remacle-Bonnet, F Garrouste, J Luis, C Rabenandrasana, J Marvaldi, G Pommier

{"title":"Surface distribution of the EGF receptor during differentiation of the human colon carcinoma cell line HT29-D4.","authors":"M Lehmann, M Remacle-Bonnet, F Garrouste, J Luis, C Rabenandrasana, J Marvaldi, G Pommier","doi":"10.3109/10799899409066040","DOIUrl":"https://doi.org/10.3109/10799899409066040","url":null,"abstract":"The clone HT29-D4 can be induced to differentiate into enterocyte-like cells, by simply removing glucose from culture medium. In this report, we used the HT29-D4 model to study the membrane segregation of the EGF receptor on epithelial intestinal cells. Differentiated and undifferentiated cells displayed a single class of EGF binding sites with similar dissociation constants. However, differentiation of HT29-D4 led to a 3-fold decrease in the total number of EGF binding sites, while the number of IGF-I binding sites was unchanged. Fifteen percent of EGF receptors present on differentiated HT29-D4 cells were localized in the apical surface, whereas 98% of IGF-I receptors were segregated to the basolateral domain. By covalent cross-linking experiments using 125I-EGF and by immunoprecipitation with an anti-EGF receptor antibody, we have characterized the HT29-D4 EGF receptor as a Mr = 165,000 protein in both differentiated and undifferentiated cells. Apical EGF receptors were functional, as evidenced by their ability to be internalized in response to EGF binding. Thus, intact and functional EGF receptors are present at the apical surface of differentiated HT29-D4 cells, suggesting the presence of EGF receptors on the apical domain of enterocytes.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 5","pages":"319-33"},"PeriodicalIF":0.0,"publicationDate":"1994-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409066040","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18817636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Independent and coordinate regulation of beta 1- and beta 2-adrenergic receptors in rat C6 glioma cells. β 1-和β 2-肾上腺素能受体在大鼠C6胶质瘤细胞中的独立和协调调节。

Journal of receptor research Pub Date : 1994-07-01 DOI: 10.3109/10799899409066037

P H Fishman, T Miller, P K Curran, G K Feussner

{"title":"Independent and coordinate regulation of beta 1- and beta 2-adrenergic receptors in rat C6 glioma cells.","authors":"P H Fishman, T Miller, P K Curran, G K Feussner","doi":"10.3109/10799899409066037","DOIUrl":"https://doi.org/10.3109/10799899409066037","url":null,"abstract":"Rat C6 glioma cells have both beta 1- and beta 2-adrenergic receptors in approximately 7:3 ratio. When the cells were exposed to the beta-adrenergic agonist isoproterenol, there was a rapid sequestration of up to 50% of the surface receptor population over a 30-min period as measured by the loss of binding of the hydrophilic ligand [3H] CGP-12177 to intact cells. Using the beta 1-selective antagonist CGP 20712A to quantify the proportion of the two subtypes, it was found that although both beta 1 and beta 2 receptors were sequestered, the latter were sequestered initially twice as fast as the former. More prolonged agonist exposure led to a down-regulation of approximately 90% of the total receptor population by 6 h as measured by the loss of binding of the more hydrophobic ligand [125I]iodocyanopindolol to cell lysates. The two subtypes, however, underwent down-regulation with similar kinetics. Treatment of the cells with agents that raise cyclic AMP levels such as cholera toxin and forskolin resulted in a slower, but still coordinated down-regulation of both subtypes. Thus, there appears to be both independent and coordinate regulation of endogenous beta 1-and beta 2-adrenergic receptors in the same cell line.","PeriodicalId":16948,"journal":{"name":"Journal of receptor research","volume":"14 5","pages":"281-96"},"PeriodicalIF":0.0,"publicationDate":"1994-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3109/10799899409066037","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18814444","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13