title: alpha-N-acetyl-beta-endorphin1-26 from the neurointermediary lobe of the rat pituitary: isolation, purification, and characterization by high-performance liquid chromatography.
abstract: The neurointermediary lobes from 190 rat pituitaries were homogenized in an acidic medium which inhibits peptidase activity and maximizes the solubilization of undamaged peptides. Octadecylsilyl-silica (ODS-silica) was used to extract the supernatant of the tissue homogenate. The ODS-silica eluate, now largely protein and salt free, was subjected to reversed-phase high-performance liquid chromatography (HPLC) employing 0.1% trifluoroacetic as counter ion. The column eluates were monitored for beta-endorphin immunoreactivity. Five immunoreactive components were observed. The most hydrophobic of these was repurified on the same HPLC column using 0.13% heptafluorobutyric acid as counter ion. Characterization of the purified peptide by gel permeation HPLC, amino acid analysis, and tryptic fragmentation indicated that it corresponded in structure to alpha-N-acetyl-beta-endorphin1-26. Amino acid analysis of the native peptide and its trypsin and carboxypeptidase fragments indicated that an alanyl residue occupies position 26. This finding is in contrast to the sequence predicted for the beta-lipotropin/corticotropin precursor by recombinant DNA techniques which suggests that the 26th residue of the beta-endorphin molecule should be valine.
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D002268:Carboxypeptidases; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004723:Endorphins / Q000302:isolation & purification*; D010455:Peptides / Q000302:isolation & purification; D010902:Pituitary Gland / Q000032:analysis; D051381:Rats; D014357:Trypsin; D001615:beta-Endorphin / Q000031:analogs & derivatives*
chemical_list: D000596:Amino Acids; D004723:Endorphins; D010455:Peptides; D001615:beta-Endorphin; C035101:N-acetyl-beta-endorphin; D002268:Carboxypeptidases; D014357:Trypsin