title: A tissue-type plasminogen activator mutant with prolonged clearance in vivo. Effect of removal of the growth factor domain.
abstract: The complete cDNA for human tissue-type plasminogen activator (t-PA) was cloned and sequenced. A mutant was constructed by using in vitro site-specific mutagenesis to delete the region encoding the growth factor domain (amino acids 51-87 inclusive). Normal and mutant t-PA species were produced using two mammalian expression systems (in human HeLa cells and mouse C127 cells). The clearance of mutant and normal t-PA from plasma was examined in vivo using a guinea pig model. Mutant t-PA derived from HeLa or C127 cells was cleared much more slowly than the cognate normal t-PA. The potential role of the growth factor domain in the recognition of t-PA by the hepatic clearance mechanism is discussed.
authors.affiliation: Beecham Pharmaceuticals Research Division, Biosciences Research Centre, Epsom, Surrey, United Kingdom.
authors.forename: M J
authors.initials: MJ
authors.lastname: Browne
authors.forename: J E
authors.initials: JE
authors.lastname: Carey
authors.forename: C G
authors.initials: CG
authors.lastname: Chapman
authors.forename: A W
authors.initials: AW
authors.lastname: Tyrrell
authors.forename: C
authors.initials: C
authors.lastname: Entwisle
authors.forename: G M
authors.initials: GM
authors.lastname: Lawrence
authors.forename: B
authors.initials: B
authors.lastname: Reavy
authors.forename: I
authors.initials: I
authors.lastname: Dodd
authors.forename: A
authors.initials: A
authors.lastname: Esmail
authors.forename: J H
authors.initials: JH
authors.lastname: Robinson
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006168:Guinea Pigs; D006801:Humans; D009154:Mutation; D013329:Structure-Activity Relationship; D010959:Tissue Plasminogen Activator / Q000097:blood / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004247:DNA; D004262:DNA Restriction Enzymes; C024404:BglII endonuclease; D015252:Deoxyribonucleases, Type II Site-Specific; D010959:Tissue Plasminogen Activator