Heterologous expression, purification, and properties of a potato protein inhibitor of serine proteinases |
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| Authors: | A S Speranskaya A A Krinitsina T A Revina N G Gerasimova Ya S Keruchen’ko A B Shevelev T A Valueva |
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| Institution: | (1) Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, 119071 Moscow, Russia;(2) Faculty of Biology, Lomonosov Moscow State University, 119992 Moscow, Russia |
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| Abstract: | The gene PKPI-B10 AF536175] encoding in potato (Solanum tuberosum L., cv. Istrinskii) a Kunitz-type protein inhibitor of proteinases (PKPI) has been cloned into the pET23a vector and then expressed in Escherichia coli. The recombinant protein PKPI-B10 obtained as inclusion bodies was denatured, separated from admixtures by ion-exchange fast protein liquid chromatography (FPLC) on MonoQ under denaturing conditions, and renatured. The native protein was additionally purified by ion-exchange FPLC on DEAE-Toyopearl. The PKPI-B10 protein effectively inhibits the activity of trypsin, significantly weaker suppresses the activity of chymotrypsin, and has no effect on other serine proteinases: human leukocyte elastase, subtilisin Carlsberg, and proteinase K, and also the plant cysteine proteinase papain. |
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| Keywords: | Kunitz-type proteinase inhibitors serine proteinases potato heterologous expression |
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