Soluble Expression and Purification of Human Tissue-type Plasminogen Activator Protease Domain 


Vol. 31,  No. 9, pp. 2607-2606, Sep.  2010
10.5012/bkcs.2010.31.9.2607


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  Abstract

Human tissue-type plasminogen activator (tPA) is a valuable thrombolytic agent used to successfully treat acute myocardial infarction, thromboembolic stroke, peripheral arterial occlusion, and venous thromboembolism. Recombinant tPA is accumulated as an inactive form in inclusion bodies of E. coli and is refolded in vitro, which is accompanied by extensive aggregation. In the present study, a tPA protease domain was expressed in an active soluble form in the cytosol of E. coli Rosetta-gami cells, which allowed disulfide bond formation and supplied the tRNA molecules required for six rarely used codons in E. coli. This strategy increased the amount of soluble protease domain protein and avoided the cumbersome refolding process. The purified protease domain not only degraded tPA substrate peptides but also formed a covalently bound complex with plasminogen activator inhibitor-1, as does full-length tPA. Soluble expression and purification of tPA domains may aid in functional analyses of this multi-domain protein, which has been implicated in many physiological and pathological processes.

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  Cite this article

[IEEE Style]

H. J. Lee and H. Im, "Soluble Expression and Purification of Human Tissue-type Plasminogen Activator Protease Domain," Bulletin of the Korean Chemical Society, vol. 31, no. 9, pp. 2607-2606, 2010. DOI: 10.5012/bkcs.2010.31.9.2607.

[ACM Style]

Hak Joo Lee and Hana Im. 2010. Soluble Expression and Purification of Human Tissue-type Plasminogen Activator Protease Domain. Bulletin of the Korean Chemical Society, 31, 9, (2010), 2607-2606. DOI: 10.5012/bkcs.2010.31.9.2607.

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ISSN(Print) 0253-2964
ISSN(Online) 1229-5949