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  • Title: Human preproendothelin-1 is converted into active endothelin-1 by baculovirus-infected insect cells.
    Author: Benatti L, Cozzi L, Zamai M, Tamburin M, Vaghi F, Caiolfa VR, Fabbrini MS, Sarmientos P.
    Journal: Biochem Biophys Res Commun; 1992 Jul 31; 186(2):753-9. PubMed ID: 1497664.
    Abstract:
    To investigate biochemical and biological parameters involved in preproendothelin-1 (preproET-1) maturation we infected Spodoptera frugiperda (Sf21) cells with a suitable engineered baculovirus vector carrying the cDNA encoding the entire human 212 amino acids precursor. Culture supernatants were tested by RIA using an anti-ET-1 serum, ET-1-like immunoreactive material (IRM) was detected in the infected Sf21 cells but not in control, wild-type or mock-infected cells. Fractionation of the culture supernatant by RP-HPLC coupled to an ET-1 specific RIA yielded two main peaks corresponding to the retention times of human bigET-1 and ET-1. Furthermore, culture supernatant of preproET-1 expressing Sf21 cells elicited a characteristic dose-response vasoconstrictive activity on rabbit vena cava, consistent with the amount of ET-1 as estimated by RP-HPLC coupled to RIA. These results suggest that insect cells possess the enzymatic activities necessary for human preproET-1 full maturation even though no such peptide has ever been found in insect cells.
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