Kosaka T, Suzuki N, Ishibashi Y, Matsumoto H, Itoh Y, Ohkubo S, Ogi K, Kitada C, Onda H, Fujino M
Discovery Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki.
J Biochem. 1994 Aug;116(2):443-9. doi: 10.1093/oxfordjournals.jbchem.a124544.
To understand the biosynthesis of the human ET family, three kinds of cloned cDNA for human prepro-endothelin-1 (prepro-ET-1), prepro-ET-2, and prepro-ET-3 were stably expressed in Chinese hamster ovary cells (CHO-K1). Immunoreactive (ir-) ET polypeptides in the culture media of the transformants were purified by reverse-phase high-performance liquid chromatography (HPLC) coupled with sandwich-type enzyme immunoassay (EIA). Amino acid sequencing and FAB mass spectrometry of the purified ir-ET polypeptides revealed the presence of human big ET-1 (1-38), big ET-2 (1-38), and big ET-3 (1-41)NH2 as intermediate forms. These results directly revealed the biosynthetic pathways of three human ETs at the peptide level.
