Isolation, Identification and Pharmacokinetic Properties of Human Tissue-Type Plasminogen Activator Species: Possible Localisation of a Clearance Recognition Site
| Autor: | Barbara Nunn, Jeff H. Robinson, Ian Dodd |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Male
medicine.medical_specialty Acylation medicine.medical_treatment Guinea Pigs Molecular Sequence Data law.invention law Internal medicine Fibrinolysis medicine Animals Amino Acid Sequence Polyacrylamide gel electrophoresis Cell Line Transformed Binding Sites Chemistry T-plasminogen activator Alternative splicing Hematology Ouchterlony double immunodiffusion Molecular biology Molecular Weight Endocrinology Sephadex Tissue Plasminogen Activator Recombinant DNA Plasminogen activator |
| Zdroj: | Thrombosis and Haemostasis. 59:523-528 |
| ISSN: | 2567-689X 0340-6245 |
| DOI: | 10.1055/s-0038-1647527 |
| Popis: | SummaryPurified preparations of recombinant tissue-type plasminogen activator (t-PA) from the recombinant Bowes melanoma cell line TRBM6 were shown to contain multiple species of plasminogen activator. Using a combination of chromatography on Sephadex G25, Sephadex G75 and Heparin Sepharose CL6B we have isolated two fibrinolytically active species, which, under nonreduced SDS PAGE, have apparent Mr = 38,000 and 56,000. Double immunodiffusion studies indicated that both species were closely related to both the t-PA B chain and t-PA itself. N-terminal sequencing identified the Mr = 38,000 species as ala160-t-PA (essentially ΔFGKI t-PA) and the Mr = 56,000 species as ser1-tyr2-gln3-glyx-cys51 t-PA (ΔF t-PA), the latter probably produced by alternative splicing of the t-PA gene. The pharmacokinetic properties of N,N dirnethy1-4-aminobenzoyl (DAB) derivatives of these activators and native t-PA were determined in the guinea pig. Whereas DAB → ΔF t-PA showed a similar, rapid plasma disappearance profile to that of DAB → t-PA, DAB → ΔFGKI t-PA was cleared significantly slower. These results suggest that a rapid clearance recognition site resides on either the growth factor or kringle 1, or both, domains of t-PA. |
| Databáze: | OpenAIRE |
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