title: Isolation of a human cDNA of urokinase and its expression in COS-1 cells.
abstract: The cDNA encoding human urokinase (UK) has been isolated from a cDNA library prepared from human normal fibroblast (WI38) cells, which had been stimulated by endothelial cell growth factor and heparin. This cDNA was sequenced and found to contain a few silent substitutions, thus encoding the same amino acids as deduced from the published genomic sequence of UK. After modification, the cDNA of UK was inserted into a transient expression vector and used to transfect COS-1 cells. The recombinant UK protein (rUK) in the serum-free medium of transfected COS-1 cells was characterized by biochemical and functional assays. These studies indicated that rUK from COS-1 cells is glycosylated, enzymatically active, and very similar to native single-chain plasminogen activator (scuPA). Therefore, such rUK can be a convenient source of scuPA for any further studies.
authors.affiliation: Biotechnology and Microbiology Division, Wyeth-Ayerst Research, Radnor, PA 19087.
authors.forename: S M
authors.initials: SM
authors.lastname: Cheng
authors.forename: S G
authors.initials: SG
authors.lastname: Lee
authors.forename: N K
authors.initials: NK
authors.lastname: Kalyan
authors.forename: W T
authors.initials: WT
authors.lastname: Hum
authors.forename: M
authors.initials: M
authors.lastname: Blume
authors.forename: R
authors.initials: R
authors.lastname: Vogel
authors.forename: P P
authors.initials: PP
authors.lastname: Hung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics*
chemical_list: D004247:DNA; D014568:Urokinase-Type Plasminogen Activator