Isolation of dermenkephalin from amphibian skin, a high-affinity delta-selective opioid heptapeptide containing a D-amino acid residue.
The predicted amino acid sequence of the biosynthetic precursor of dermorphin, a highly potent and nearly specific mu-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 micrograms/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 delta-opioid receptors. Because of its origin and its delta-opioid (enkephalin) activity and specificity, this novel D-amino acid containing peptide is named dermenkephalin.
Mor
A
A
Laboratoire de Bioactivation des Peptides, Institut Jacques Monod, Université Paris, France.
Delfour
A
A
Sagan
S
S
Amiche
M
M
Pradelles
P
P
Rossier
J
J
Nicolas
P
P
D000110:Acetylcholinesterase; D000595:Amino Acid Sequence; D000663:Amphibians; D000818:Animals; D001921:Brain / Q000378:metabolism; D003429:Cross Reactions; D004557:Electric Organ / Q000201:enzymology; D004593:Electrophorus; D007118:Immunoassay; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D011957:Receptors, Opioid / Q000378:metabolism*; D017465:Receptors, Opioid, delta; D012867:Skin / Q000032:analysis*
D009842:Oligopeptides; D011957:Receptors, Opioid; D017465:Receptors, Opioid, delta; C059567:deltorphin; D000110:Acetylcholinesterase