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Recombinant alkaline serine protease II degrades scrapie isoform of prion protein |
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| Authors: | Zhao Hui Kazuhisa Minamiguchi Hiroyasu Doi Naoko Kinoshita Hiroaki Kanouchi Tatsuzo Oka |
| Affiliation: | (1) Department of Veterinary Physiology, Faculty of Agriculture, Kagoshima University, 890-0065 Kagoshima, Japan;(2) Hanno Research Center, Taiho Pharmaceutical Co. Ltd., 1-27, Misugidai, 357-8527 Hanno, Saitama, Japan;(3) Microbial Chemistry Research Foundation, Shinagawa, 141-0021 Tokyo, Japan;(4) Department of Biochemistry, Kawasaki Medical School, Kurasiki, 701-0192 Okayama, Japan |
| Abstract: | Summary An efficient Escherichia coli expression system for the production of mature-type alkaline serine protease II (mASP II) has been constructed. Complementary deoxyribonucleic acid-encoding mASP II was inserted into the inducible bacterial expression vector pGE-30. After introduction into E, coli, the plasmid was expressed by isopropyl-1-thio-β-d-galactopyranoside, and the recombinant product was purified using a Ni-nitrilotriacetic acid column The purified product had the expected NH2-terminal sequence and showed a scrapie isoform of prion protein-degrading activity using hamster scrapie 263K prions as a substrate. |
| Keywords: | alkaline serine protease perchloric acid-soluble protein Escherichia coli expression His tag |
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