期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1979
卷号：76
期号：3
页码：1448-1452
DOI：10.1073/pnas.76.3.1448
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Enzymatically dispersed bovine hypothalamic or cortical tissue was maintained in culture in the presence of 3H-labeled amino acids. After such incubation, extracts of cells and of media contained 3H-labeled products that were specifically bound by immobilized affinity-purified antisera to corticotropin (ACTH) and {beta}-endorphin. The majority of these products eluted in the void volume (V0) upon Sephadex G-50 gel filtration; minor 3H-labeled products eluted in the regions of the ovine {beta}-lipotropin marker and in fractions having apparent molecular weights of approximately 12,000 and 3800. Sequential use of these immobilized antisera revealed that most of the V0 material contained both ACTH and {beta}-endorphin antigenic determinants within the same molecule(s), whereas retarded material contained only one of the determinants. When this V0 material was rerun on a Sephadex G-75 column, it coeluted with the 31-kilodalton precursor of both ACTH and {beta}-endorphin obtained from a bovine anterior pituitary extract. Thus, the high molecular weight immunoreactive ACTH/{beta}-endorphin-like 3H-labeled product(s) derived from the hypothalamic culture is similar to the pituitary-derived precursor in containing the dual antigenic determinants and in its gel filtration characteristics. In contrast, the cortex-derived cell preparation was devoid of 3H-labeled products specifically reactive with the antisera employed.
