Synthesis and binding characteristics of two sulfhydryl‐reactive probes for vasopressin receptors

FEBS Letters

Pávó, Imre¹  Kojro, Elzbieta¹  Fahrenholz, Falk¹ 

[1] Max Planck Institut für Biophysik, Kennedyallee 70, W-6000 Frankfurt am Main 70, Germany

关键词: Vasopressin;    Vasopressin V2 receptor;    Sulfhydryl reagent;    ACN;    acetonitrile;    AVP;    [8-arginine]vasopressin;    BOC;    tert-butyloxycarbonyl;    Dab;    2;    3-diaminobutyric acid;    DCC;    dicyclohexylcarbodiimide;    DMF;    N;    N-dimethylformamide;    EPPS;    N-(2-hydroxyethyl)-piperazine-N′-(3-propanesulfonic acid);    NEM;    N-ethylmalcimide;    TFA;    trifluoroacetic acid.;   

DOI  :  10.1016/0014-5793(93)81736-J

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The present study describes the synthesis and receptor binding affinities of the sulfhydryl-reactive vasopressin analogs deamino[Dab(N ⁸-N-maleoyl-β-alanin e)⁴]AVP (1a) and deamino[Lys(N ⁸-N-maleoyl-β-alanine)⁸VP (2a). The analogs were obtained by introducing the sulfhydryl-reactive maleoyl-β-alanyl group at the δ-amino group of Dab⁴ in deamino[Dab⁴]IAVP (1) and at the ε-amino group of Lys⁸ in deamino[Lys⁸]VP (2), which were synthesized by the solid-phase method. Furthermore, the analog modified at Dab⁴ was prepared as tritium labeled compound (1b) after catalytic iodine tritium exchange at Tyr² in deamino[Dab⁴]AVP. The sulfhydryl-reactive vasopressin analogs retained high binding affinity for the V₂ vasopressin receptor in membranes derived from bovine kidney inner medulla. Apparent dissociation constants K _d of 45 nM (compound 1a) and 15 nM (compound 2a) were determined. Incubation of the ligand receptor complexes at pH 5.5 resulted in dissociation of the sulfhydryl-reactive vasopressin analogs from the V₂ receptor. No indications of a covalent reaction between analogs 1a, 2a and 1b and sulfhydryl groups in or close to the hormone binding site of the V₂ receptor were found.

【 授权许可】

Unknown