A tissue-type plasminogen activator mutant with prolonged clearance in vivo. Effect of removal of the growth factor domain

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...

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Bibliographic Details
Published in:The Journal of biological chemistry 1988-02, Vol.263 (4), p.1599-1602
Main Authors: Browne, M J, Carey, J E, Chapman, C G, Tyrrell, A W, Entwisle, C, Lawrence, G M, Reavy, B, Dodd, I, Esmail, A, Robinson, J H
Format: Article
Language:English
Subjects:
Animals
Bacterial Proteins
Base Sequence
Cloning, Molecular
Deoxyribonucleases, Type II Site-Specific
DNA - analysis
DNA Restriction Enzymes - metabolism
Electrophoresis, Polyacrylamide Gel
Gene Expression Regulation
Guinea Pigs
Humans
Mutation
Structure-Activity Relationship
Tissue Plasminogen Activator - blood
Tissue Plasminogen Activator - genetics
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)77918-6