PMID: 6580616Nov 1, 1983Paper

Latent tissue plasminogen activator produced by human endothelial cells in culture: evidence for an enzyme-inhibitor complex

Proceedings of the National Academy of Sciences of the United States of America
E G Levin

Abstract

Conditioned medium from cultures of human umbilical vein endothelial cells was analyzed for the presence of tissue plasminogen activator (tPA) and urokinase. Immunoprecipitation studies using metabolically labeled conditioned medium and anti-tPA IgG revealed a single band on autoradiographs corresponding to a Mr of 100,000. No bands were observed after immunoprecipitation with anti-urokinase IgG. The Mr 100,000 tPA was found to be inactive and did not bind to fibrin clots. However, exposure of this tPA to 1% NaDodSO4 resulted in the appearance of plasminogen activator activity with no apparent change in its Mr. Treatment with 10 mM diisopropylfluorophosphate prior to NaDodSO4 activation did not inhibit the NaDodSO4-induced appearance of plasminogen activator activity, indicating that the active site was not available for diisopropylfluorophosphate binding. The possibility that the properties of this Mr 100,000 tPA reflected a tPA-inhibitor complex was examined. Attempts to dissociate such a complex by denaturation, reduction, or extremes of temperature were not successful. However, after treatment of conditioned medium with 1 M hydroxylamine in the presence of 0.1% NaDodSO4, the Mr of the anti-tPA immunoprecipitable material de...Continue Reading

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Citations

Oct 1, 1991·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·G LeloupG Vaes
Jul 1, 1994·Transplant International : Official Journal of the European Society for Organ Transplantation·M ShimadaK Sugimachi
Jan 1, 1987·Archives of Dermatological Research·T Hunziker, J D Vassalli
Mar 1, 1989·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·D Dichek, T Quertermous
Nov 4, 2008·Molecular and Cellular Biochemistry·Moyuru HayashiMotoyuki Shimonaka
Apr 1, 1989·Developmental Biology·S Pino-Heiss, G Schubiger
Jan 1, 1987·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·G K Scott
Mar 1, 1993·Trends in Cardiovascular Medicine·S W LeeD A Dichek
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·B E SobelS J Sarnoff
Dec 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·L A EricksonD J Loskutoff
Feb 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·Y SakataD J Loskutoff
Aug 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·E F PlowM H Ginsberg
Jan 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·D HantaïB W Festoff
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Jul 1, 1984·The Journal of Clinical Investigation·R D Rosenberg, J S Rosenberg
Dec 1, 1984·The Journal of Clinical Investigation·E G LevinL A Harker

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