Identification of beta-endorphins in the pituitary gland and blood plasma of the common carp (Cyprinus carpio).
Carp beta-endorphin is posttranslationally modified by N-terminal acetylation and C-terminal cleavage. These processes determine the biological activity of the beta-endorphins. Forms of beta-endorphin were identified in the pars intermedia and the pars distalis of the pituitary gland of the common carp (Cyprinus carpio), as well as the forms released in vitro and into the blood. After separation and quantitation by high performance liquid chromatography (HPLC) coupled with radioimmunoassay, the beta-endorphin immunoreactive products were identified by electrospray ionisation mass spectrometry and peptide sequencing. The release of beta-endorphins by the pituitary gland was studied after stimulation with corticotrophin-releasing factor (CRF) in vitro. In the pars intermedia, eight N-acetylated truncated forms were identified. Full length N-acetyl beta-endorphin(1-33) coeluted with N-acetyl beta-endorphin(1-29) and these forms together amounted to over 50% of total immunoreactivity. These products were partially processed to N-acetyl betaendorphin(1-15) (30.8% of total immunoreactivity) and N-acetyl beta-endorphin(1-10) (3.1%) via two different cleavage pathways. The acetylated carp homologues of mammalian alpha- and gamma-endorphin were also found. N-acetyl beta-endorphin(1-15) and (1-29) and/or (1-33) were the major products to be released in vitro, and were the only acetylated beta-endorphins found in blood plasma, although never together. CRF stimulated the release of opioid beta-endorphin from the pars distalis. This non-acetylated beta-endorphin represents the full length peptide and is the most abundant form in plasma.
van Den Burg
E H
EH
Department of Animal Physiology, Faculty of Science, University of Nijmegen, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands.
Metz
J R
JR
Arends
R J
RJ
Devreese
B
B
Vandenberghe
I
I
Van Beeumen
J
J
Wendelaar Bonga
S E
SE
Flik
G
G
D000818:Animals; D002347:Carps / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D008970:Molecular Weight; D010902:Pituitary Gland / Q000033:anatomy & histology / Q000737:chemistry* / Q000378:metabolism; D011863:Radioimmunoassay; D021241:Spectrometry, Mass, Electrospray Ionization; D013268:Stimulation, Chemical; D001615:beta-Endorphin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood
D001615:beta-Endorphin; C035101:N-acetyl-beta-endorphin; D003346:Corticotropin-Releasing Hormone