Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells
Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Wehmeyer, Andrea [verfasserIn] Schulz, Rüdiger [verfasserIn] |
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| Format: |
E-Artikel |
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| Erschienen: |
Oxford, UK: Blackwell Science Ltd ; 1997 |
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| Schlagwörter: |
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| Umfang: |
Online-Ressource |
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| Reproduktion: |
2002 ; Blackwell Publishing Journal Backfiles 1879-2005 |
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| Übergeordnetes Werk: |
In: Journal of neurochemistry - Oxford : Wiley-Blackwell, 1956, 68(1997), 4, Seite 0 |
| Übergeordnetes Werk: |
volume:68 ; year:1997 ; number:4 ; pages:0 |
| Links: |
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| DOI / URN: |
10.1046/j.1471-4159.1997.68041361.x |
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NLEJ243168934 |
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| 520 | |a Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. | ||
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10.1046/j.1471-4159.1997.68041361.x doi (DE-627)NLEJ243168934 DE-627 ger DE-627 rakwb Wehmeyer, Andrea verfasserin aut Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells Oxford, UK Blackwell Science Ltd 1997 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. 2002 Blackwell Publishing Journal Backfiles 1879-2005 |2002|||||||||| Baculovirus Schulz, Rüdiger verfasserin aut In Journal of neurochemistry Oxford : Wiley-Blackwell, 1956 68(1997), 4, Seite 0 Online-Ressource (DE-627)NLEJ243927584 (DE-600)2020528-4 1471-4159 nnns volume:68 year:1997 number:4 pages:0 http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 68 1997 4 0 |
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10.1046/j.1471-4159.1997.68041361.x doi (DE-627)NLEJ243168934 DE-627 ger DE-627 rakwb Wehmeyer, Andrea verfasserin aut Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells Oxford, UK Blackwell Science Ltd 1997 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. 2002 Blackwell Publishing Journal Backfiles 1879-2005 |2002|||||||||| Baculovirus Schulz, Rüdiger verfasserin aut In Journal of neurochemistry Oxford : Wiley-Blackwell, 1956 68(1997), 4, Seite 0 Online-Ressource (DE-627)NLEJ243927584 (DE-600)2020528-4 1471-4159 nnns volume:68 year:1997 number:4 pages:0 http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 68 1997 4 0 |
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10.1046/j.1471-4159.1997.68041361.x doi (DE-627)NLEJ243168934 DE-627 ger DE-627 rakwb Wehmeyer, Andrea verfasserin aut Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells Oxford, UK Blackwell Science Ltd 1997 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. 2002 Blackwell Publishing Journal Backfiles 1879-2005 |2002|||||||||| Baculovirus Schulz, Rüdiger verfasserin aut In Journal of neurochemistry Oxford : Wiley-Blackwell, 1956 68(1997), 4, Seite 0 Online-Ressource (DE-627)NLEJ243927584 (DE-600)2020528-4 1471-4159 nnns volume:68 year:1997 number:4 pages:0 http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 68 1997 4 0 |
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10.1046/j.1471-4159.1997.68041361.x doi (DE-627)NLEJ243168934 DE-627 ger DE-627 rakwb Wehmeyer, Andrea verfasserin aut Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells Oxford, UK Blackwell Science Ltd 1997 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. 2002 Blackwell Publishing Journal Backfiles 1879-2005 |2002|||||||||| Baculovirus Schulz, Rüdiger verfasserin aut In Journal of neurochemistry Oxford : Wiley-Blackwell, 1956 68(1997), 4, Seite 0 Online-Ressource (DE-627)NLEJ243927584 (DE-600)2020528-4 1471-4159 nnns volume:68 year:1997 number:4 pages:0 http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 68 1997 4 0 |
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10.1046/j.1471-4159.1997.68041361.x doi (DE-627)NLEJ243168934 DE-627 ger DE-627 rakwb Wehmeyer, Andrea verfasserin aut Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells Oxford, UK Blackwell Science Ltd 1997 Online-Ressource nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. 2002 Blackwell Publishing Journal Backfiles 1879-2005 |2002|||||||||| Baculovirus Schulz, Rüdiger verfasserin aut In Journal of neurochemistry Oxford : Wiley-Blackwell, 1956 68(1997), 4, Seite 0 Online-Ressource (DE-627)NLEJ243927584 (DE-600)2020528-4 1471-4159 nnns volume:68 year:1997 number:4 pages:0 http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x text/html Verlag Deutschlandweit zugänglich Volltext GBV_USEFLAG_U ZDB-1-DJB GBV_NL_ARTICLE AR 68 1997 4 0 |
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Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells |
| abstract |
Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. |
| abstractGer |
Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. |
| abstract_unstemmed |
Abstract: “High 5” cells derived from Trichoplusia ni ovaries were infected with baculovirus bearing the cDNA of the mouse δ-opioid receptor. The maximal binding capacity for the narcotic antagonist [3H]naltrindole was 1.4 pmol/mg of membrane protein, and that for the agonist [3H][d-penicillamine2,d-penicillamine5]enkephalin (DPDPE) was 0.3 pmol/mg. DPDPE proved highly potent in competing with its tritiated analogue at δ-receptors of NG108-15 hybrid cells and of High 5 and Sf9 insect cells. However, in insect cells the opioid was more than 100-fold less effective in competing with [3H]naltrindole as compared with the mammalian cells. This decline in potency was counteracted in a dose-dependent manner by exposure of High 5 membranes to the exogenous G protein Go, which increased the binding capacity for DPDPE. Functional studies revealed a dose-dependent inhibition (up to 30%) by opioids on forskolin-stimulated cyclic AMP synthesis, and this effect was potentiated by Go. Quantification of Gαo and Gαi disclosed striking differences between Sf9 and High 5 insect cells, both of which overexpressed the cloned δ-opioid receptor. Although no inhibitory G proteins were detected in membranes of Sf9 cells, High 5 cells contained 0.5 pmol of Gαo/mg of membrane protein, and a 20-fold higher concentration for Gαi. The distinct G-protein expression in insect cells may be considered an advantage for studying functions of G protein-coupled receptors. |
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| title_short |
Overexpression of δ-Opioid Receptors in Recombinant Baculovirus-Infected Trichoplusia ni“High 5” Insect Cells |
| url |
http://dx.doi.org/10.1046/j.1471-4159.1997.68041361.x |
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Schulz, Rüdiger |
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| doi_str |
10.1046/j.1471-4159.1997.68041361.x |
| up_date |
2024-07-06T04:33:24.992Z |
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