Isolation of a human cDNA of urokinase and its expression in COS-1 cells.
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.
Cheng
S M
SM
Biotechnology and Microbiology Division, Wyeth-Ayerst Research, Radnor, PA 19087.
Lee
S G
SG
Kalyan
N K
NK
Hum
W T
WT
Blume
M
M
Vogel
R
R
Hung
P P
PP
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*
D004247:DNA; D014568:Urokinase-Type Plasminogen Activator