Further analysis of post-translational processing of beta-endorphin in rat intermediate pituitary.

Rat intermediate pituitary cells maintained in culture synthesize the same forms of beta-endorphin observed in intermediate pituitary extracts. Biosynthetically labeled intermediate pituitary beta-endorphin-sized material was fractionated by ion exchange chromatography on sulfopropyl-Sephadex and the identities of the major peaks were determined by co-chromatography with synthetic marker peptides, gel filtration, and analysis of pronase, chymotrypsin, and trypsin digests. Peaks of alpha-N-acetyl-beta-endorphin(1-27), alpha-N-acetyl-beta-endorphin(1-31), and beta-endorphin(1-31) were identified and a fourth peak (eluting from the sulfopropyl-Sephadex column at 0.18 M NaCl) was tentatively identified as alpha-N-acetyl-beta-endorphin(1-26). Analysis of beta-endorphin synthesized in the presence of [35S]methionine and [3H]histidine confirmed the absence of His in the material eluting at 0.18 M NaCl. Based on both steady labeling and pulse-chase incubations, beta-endorphin(1-31) was the first form of labeled beta-endorphin-sized material to appear in cell extracts. This molecule was quickly N-acetylated on its NH2-terminal tyrosine residue and was then more slowly converted to alpha-N-acetyl-beta-endorphin(1-27) and then to alpha-N-acetyl-beta-endorphin(1-26).