Expression of human preproendothelin-1 cDNA in COS cells results in the production of mature vasoactive endothelin-1.
Transient transfection of simian kidney (COS) cells with a recombinant plasmid encoding human preproendothelin-1 resulted in the production of biologically active endothelin-1. Conditioned medium from human preproendothelin-1 transfected cells demonstrated a significant increase in immunoreactive endothelin and big endothelin which co-eluted, when analyzed by reverse phase HPLC, with synthetic endothelin-1 and big endothelin-1, respectively. In addition, biological activity was confirmed by both inhibition of [125I]endothelin-1 binding to rat cerebellar and renal medullary membrane endothelin receptors and in vitro vasoconstriction of rabbit aorta. This is the first demonstration that human preproendothelin-1 is capable of being processed to a vasoactive form in a heterologous system and suggests that human preproendothelin-1 transfected COS cells may provide a useful model system for the study of endothelin biosynthesis.
DiLella
A G
AG
Department of Molecular Genetics, SmithKline Beecham Pharmaceuticals King of Prussia, PA 19406.
Ohlstein
E
E
Elshourbagy
N
N
Bhatnagar
P K
PK
Nambi
P
P
DeWolf
W E
WE
Caltabiano
M M
MM
D000818:Animals; D001681:Biological Assay; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D016232:Endothelins / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D011817:Rabbits; D011994:Recombinant Proteins; D014162:Transfection; D014661:Vasoconstriction
D016232:Endothelins; D011994:Recombinant Proteins; D004247:DNA