Isolation, Identification and Pharmacokinetic Properties of Human Tissue-Type Plasminogen Activator Species: Possible Localisation of a Clearance Recognition Site

Ian Dodd; Barbara Nunn; Jeff H Robinson

doi:10.1055/s-0038-1647527

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1988; 59(03): 523-528
DOI: 10.1055/s-0038-1647527

Original Article

Schattauer GmbH Stuttgart

Isolation, Identification and Pharmacokinetic Properties of Human Tissue-Type Plasminogen Activator Species: Possible Localisation of a Clearance Recognition Site

Ian Dodd

The Beecham Pharmaceuticals Research Division, Biosciences Research Centre, Great Burgh, Epsom, Surrey, UK

,

Barbara Nunn

The Beecham Pharmaceuticals Research Division, Biosciences Research Centre, Great Burgh, Epsom, Surrey, UK

,

Jeff H Robinson

The Beecham Pharmaceuticals Research Division, Biosciences Research Centre, Great Burgh, Epsom, Surrey, UK

› Author Affiliations

Abstract

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Summary

Purified 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 M_r = 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 M_r = 38,000 species as ala₁₆₀-t-PA (essentially ΔFGKI t-PA) and the M_r = 56,000 species as ser₁-tyr₂-gln₃-gly_x-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.

Keywords

Tissue-type plasminogen activator - Fibrinoly - Pharmacokinetics

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References

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