DIFFERENTIAL MECHANISMS FOR THE N-ACETYLATION OF ALPHA-MELANOCYTE-STIMULATING HORMONE AND BETA-ENDORPHIN IN THE INTERMEDIATE PITUITARY OF THE FROG, XENOPUS-LAEVIS

Immunohistochemical analysis of the pituitary of Xenopus laevis revealed the colocalization of alpha-melanocyte-stimulating-hormone (MSH)-related immunoreactivity and N-acetyl-beta-endorphin-related immunoreactivity in the cells of the intermediate pituitary. In order to determine whether the immunoreactive N-acetylated beta-endorphin is released in parallel with the immunoreactive alpha-MSH, intermediate pituitaries were incubated in L-15 medium for 24 h. The medium and an acid extract of the intermediate pituitaries from each incubation were separately fractionated by a combination of gel filtration chromatography, reverse-phase high-performance liquid chromatography, and cation exchange chromatography. In the intermediate pituitary extract, the major form of alpha-MSH had chromatographic properties which corresponded to nonacetylated alpha-MSH (ACTH)(1-13)amide;whereas the major form of beta-endorphin had an apparent molecular weight of 1.2 kDa and was N-acetylated. The 1.2-kDa form of beta-endorphin and ACTH(1-13)amide were present in equimolar amounts. Analysis of the medium indicated that both end products were released in parallel. However, as reported in the literature, there was a significant increase in the N-acetylation of ACTH(1-13)amide during secretion. There was no further processing of beta-endorphin during secretion. Collectively, these observations indicate that in the intermediate pituitary of X. laevis there are separate mechanisms for the N-acetylation of alpha-MSH and beta-endorphin.