Isolation of dermenkephalin from amphibian skin, a high‐affinity (δ‐selective opioid heptapeptide containing a D‐amino acid residue

FEBS Letters

Pradelles, Philippe³  Mor, Amram¹  Nicolas, Pierre¹  Sagan, Sandrine¹  Rossier, Jean²  Amiche, Mohamed¹  Delfour, Antoine¹ 

[1] Laboratoire de Bioactivation des Peptides, Institut Jacques Monod, Université Paris 7, 2 place Jussieu, 75251 Paris Cedex 05, France;Laboratoire de Physiologie Nerveuse, CNRS, 91198 Gif-sur-Yvette Cedex, France;Section de Pharmacologie et d'Immunologie, Département de Biologie, Centre d'Etudes Nucléaires, Saclay, 91191 Gif-sur-Yvette Cedex, France

关键词: Opioid receptor;    δ-;    Amino acid containing peptide;    D-;    Dermorphin;    Dermenkephalin;    DREK;    dermenkephalin;    DRM;    dermorphin;    EIA;    enzyme immunoassay;    BSA;    bovine serum albumin;    RP-HPLC;    reverse-phase high-performance liquid chromatography;    DTLET;    [D-Thr2;    Leu5]-enkephalyl-Thr6;    NHS;    N-hydroxysuccinimide;    DCC;    N;    N′-dicyclohexylcarbodiimide;   

DOI  :  10.1016/0014-5793(89)81104-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The predicted amino acid sequence of the biosynthetic precursor of dermorphin, a highly potent and nearly specific μ-opioid peptide from amphibian skin, contains four repeats of the dermorphin progenitor sequence and one single copy of a different heptapeptide sequence. We have developed a specific enzyme immunoassay and used synthetic peptides to detect and purify the new predicted heptapeptide 2.4 μg/g dry skin) from the skin of the Phyllomedusa sauvagei frog from which dermorphin was originally isolated. The identity of the novel pro-dermorphin related peptide, Tyr-D-Met-Phe-His-Leu-Met-Asp-NH₂, was established by co-chromatography with synthetic peptides on reverse-phase HPLC, immunological analysis, gas-phase sequencing, mass spectrometry and by pharmacological assays. Opioid-binding assays in vitro demonstrated that both the natural and synthetic heptapeptides displayed exceptionally high selectivity and affinity towards the δ-opioid receptors. Because of its origin and its δ-opioid (enkephalin) activity and specificity, this novel D-amino acid containing peptide is named dermenkephalin.

【 授权许可】

Unknown