Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells

Ivone Gomes; Achla Gupta; S. P. Singh; Shail K. Sharma

doi:10.1016/S0014-5793(99)00878-9

Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells

Ivone Gomes, Achla Gupta, S. P. Singh, Shail K. Sharma

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

Monoclonal antibodies generated against multiple antigenic peptides of the N-terminal sequence (³LVPSARAELQSSPLV¹⁷) of the cloned δ opioid receptor immunoprecipitated a 58 kDa protein from CHAPS-solubilized NG108-15 membranes. The immunoprecipitates bound [³H]DPDPE - but not [³H]DAMGO - with a K(d) of 6.4 nM and a B(max) of 75 pM. Western blot analysis revealed a distinct band of 58 kDa. The antibodies inhibited basal and PGE₁-stimulated cAMP levels, and mimicked the effect of agonists manifest in a compensatory increase in cAMP formation. The antibody will be potentially useful in the analysis of functional epitopes on the δ opioid receptor. Copyright (C) 1999 Federation of European Biochemical Societies.

Original language	English
Pages (from-to)	126-130
Number of pages	5
Journal	FEBS Letters
Volume	456
Issue number	1
DOIs	https://doi.org/10.1016/S0014-5793(99)00878-9
State	Published - 30 Jul 1999
Externally published	Yes

Keywords

Adenylate cyclase
Antibody
G protein
Opioid receptor
Signal transduction
cAMP

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10.1016/S0014-5793(99)00878-9

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@article{095995b917f54a05af88f60320df5d94,

title = "Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells",

abstract = "Monoclonal antibodies generated against multiple antigenic peptides of the N-terminal sequence (3LVPSARAELQSSPLV17) of the cloned δ opioid receptor immunoprecipitated a 58 kDa protein from CHAPS-solubilized NG108-15 membranes. The immunoprecipitates bound [3H]DPDPE - but not [3H]DAMGO - with a K(d) of 6.4 nM and a B(max) of 75 pM. Western blot analysis revealed a distinct band of 58 kDa. The antibodies inhibited basal and PGE1-stimulated cAMP levels, and mimicked the effect of agonists manifest in a compensatory increase in cAMP formation. The antibody will be potentially useful in the analysis of functional epitopes on the δ opioid receptor. Copyright (C) 1999 Federation of European Biochemical Societies.",

keywords = "Adenylate cyclase, Antibody, G protein, Opioid receptor, Signal transduction, cAMP",

author = "Ivone Gomes and Achla Gupta and Singh, {S. P.} and Sharma, {Shail K.}",

note = "Funding Information: This work is supported by CSIR Grant 21(0397)/97/EMRII and DBT Grant BT/R and D/15/26/93 to S.K.S.",

year = "1999",

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doi = "10.1016/S0014-5793(99)00878-9",

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pages = "126--130",

journal = "FEBS Letters",

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T1 - Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells

AU - Gomes, Ivone

AU - Gupta, Achla

AU - Singh, S. P.

AU - Sharma, Shail K.

N1 - Funding Information: This work is supported by CSIR Grant 21(0397)/97/EMRII and DBT Grant BT/R and D/15/26/93 to S.K.S.

PY - 1999/7/30

Y1 - 1999/7/30

N2 - Monoclonal antibodies generated against multiple antigenic peptides of the N-terminal sequence (3LVPSARAELQSSPLV17) of the cloned δ opioid receptor immunoprecipitated a 58 kDa protein from CHAPS-solubilized NG108-15 membranes. The immunoprecipitates bound [3H]DPDPE - but not [3H]DAMGO - with a K(d) of 6.4 nM and a B(max) of 75 pM. Western blot analysis revealed a distinct band of 58 kDa. The antibodies inhibited basal and PGE1-stimulated cAMP levels, and mimicked the effect of agonists manifest in a compensatory increase in cAMP formation. The antibody will be potentially useful in the analysis of functional epitopes on the δ opioid receptor. Copyright (C) 1999 Federation of European Biochemical Societies.

AB - Monoclonal antibodies generated against multiple antigenic peptides of the N-terminal sequence (3LVPSARAELQSSPLV17) of the cloned δ opioid receptor immunoprecipitated a 58 kDa protein from CHAPS-solubilized NG108-15 membranes. The immunoprecipitates bound [3H]DPDPE - but not [3H]DAMGO - with a K(d) of 6.4 nM and a B(max) of 75 pM. Western blot analysis revealed a distinct band of 58 kDa. The antibodies inhibited basal and PGE1-stimulated cAMP levels, and mimicked the effect of agonists manifest in a compensatory increase in cAMP formation. The antibody will be potentially useful in the analysis of functional epitopes on the δ opioid receptor. Copyright (C) 1999 Federation of European Biochemical Societies.

KW - Adenylate cyclase

KW - Antibody

KW - G protein

KW - Opioid receptor

KW - Signal transduction

KW - cAMP

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U2 - 10.1016/S0014-5793(99)00878-9

DO - 10.1016/S0014-5793(99)00878-9

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C2 - 10452543

AN - SCOPUS:0032775853

SN - 0014-5793

VL - 456

SP - 126

EP - 130

JO - FEBS Letters

JF - FEBS Letters

IS - 1

ER -

Monoclonal antibody to the δ opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells

Abstract

Keywords

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