Isolation of immunoreactive beta-endorphin-related and Met-enkephalin-related peptides from the posterior pituitary of the amphibian, Xenopus laevis.
Acid extracts of the posterior pituitary of the amphibian, Xenopus laevis, were analyzed with two heterologous region specific beta-endorphin RIAs. Following gel filtration chromatography and cation exchange chromatography four peaks of immunoreactivity were detected. All four peaks were detected with a N-acetyl specific beta-endorphin RIA. Peak I represented 92% of the total immunoreactivity isolated following cation exchange chromatography. This peak had a net positive charge at pH 2.5 of +1 and an apparent molecular weight of 1.4 Kd. Following reverse phase HPLC, Peak I fractionated into two peaks: Peak Ia and Peak Ib. Both peaks were detected with the N-acetyl specific beta-endorphin RIA and a Met-enkephalin RIA, however, neither peak co-migrated with either Met-enkephalin or N-acetyl-beta-endorphin(1-16). At present it is not clear whether Peak I is derived from pro-opiomelanocortin or one of the other opioid polyproteins. Peaks II, III, and IV represented 8% of the total immunoreactivity recovered following cation exchange chromatography. These peaks had net positive charges of +3, +4, and +5, respectively, and apparent molecular weights of 2.8, 3.2, and 3.5 Kd, respectively. These apparently N-acetylated beta-endorphin-sized forms are minor end products of the pro-opiomelanocortin biosynthetic pathway.
Dores
R M
RM
University of Denver, Department of Biological Sciences, CO 80208.
Rothenberg
M E
ME
D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004744:Enkephalin, Methionine / Q000276:immunology / Q000302:isolation & purification*; D008970:Molecular Weight; D010904:Pituitary Gland, Posterior / Q000032:analysis*; D011863:Radioimmunoassay; D014982:Xenopus laevis; D001615:beta-Endorphin / Q000276:immunology / Q000302:isolation & purification*
D004744:Enkephalin, Methionine; D001615:beta-Endorphin