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

FEBS Letters

Singh, S.P¹  Gomes, Ivone¹  Gupta, Achla¹  Sharma, Shail K¹ 

[1] Department of Biochemistry, All India Institute of Medical Sciences, New Delhi 110029, India

关键词: Adenylate cyclase;    Antibody;    cAMP;    G protein;    Opioid receptor;    Signal transduction;    cAMP;    cyclic adenosine-3′;    5′-monophosphate;    CHAPS;    3-[(3-cholamidopropyl) dimethyl ammonio] 1-propane-sulfonate;    DAMGO;    [D-Ala2;    N-methyl-Phe4;    Gly-ol] enkephalin;    DPDPE;    D-Pen2;    D-Pen5-enkephalin;    DMEM;    Dulbecco's modified Eagle's medium;    HEPES;    N-[2-hydroxyethy1]piperazine-N′-[2-ethanesulfonic acid];    HRPO;    horseradish peroxidase;    K d;    equilibrium dissociation constant;    MAP;    multiple antigenic peptide;    Nt-Ab;    N-terminal monoclonal antibody;    PBS;    phosphate-buffered saline;    PGE1;    prostaglandin E1;    PMSF;    phenylmethylsulfonyl fluoride;    SDS;    sodium dodecyl sulfate;    TBS;    Tris-buffered saline;    TTBS;    Tris-buffered saline with Tween-20 (0.1%);   

DOI  :  10.1016/S0014-5793(99)00878-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown