Human preproendothelin-1 is converted into active endothelin-1 by baculovirus-infected insect cells.
Biochem Biophys Res Commun
; 186(2): 753-9, 1992 Jul 31.
Article
em En
| MEDLINE
| ID: mdl-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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Base de dados:
MEDLINE
Assunto principal:
Precursores de Proteínas
/
Baculoviridae
/
Endotelinas
/
Vetores Genéticos
Idioma:
En
Ano de publicação:
1992
Tipo de documento:
Article