Journal of Microbiology and Biotechnology

  • 제28권10호
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  • Pages.1749-1759
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  • 2018
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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High-Level Production of High-Purity Human and Murine Recombinant Prion Proteins Functionally Compatible to In Vitro Seeding Assay

  • Hwang, Hae-Gwang (Department of Pharmacy, Sunchon National University) ;
  • Kim, Dae-Hwan (Department of Pharmacy and Institute of Pharmaceutical Science & Technology, Hanyang University) ;
  • Lee, Jeongmin (Division of Zoonoses, Center for Immunology and Pathology, National Institute of Health, Korea Centers for Disease Control and Prevention) ;
  • Mo, Youngwon (Department of Pharmacy and Institute of Pharmaceutical Science & Technology, Hanyang University) ;
  • Lee, Se-Hoon (Department of Pharmacy, Sunchon National University) ;
  • Lee, Yongjin (Department of Pharmacy, Sunchon National University) ;
  • Hyeon, Jae Wook (Division of Zoonoses, Center for Immunology and Pathology, National Institute of Health, Korea Centers for Disease Control and Prevention) ;
  • Lee, Sol Moe (Division of Zoonoses, Center for Immunology and Pathology, National Institute of Health, Korea Centers for Disease Control and Prevention) ;
  • Cheon, Yong-Pil (School of Biological Sciences and Chemistry, Sungshin Women's University) ;
  • Choi, Eun-Kyoung (Ilsong Institute of Life Science, Hallym University) ;
  • Kim, Su Yeon (Division of Zoonoses, Center for Immunology and Pathology, National Institute of Health, Korea Centers for Disease Control and Prevention) ;
  • Lee, Yeong Seon (Division of Zoonoses, Center for Immunology and Pathology, National Institute of Health, Korea Centers for Disease Control and Prevention) ;
  • Son, Young-Jin (Department of Pharmacy, Sunchon National University) ;
  • Ryou, Chongsuk (Department of Pharmacy and Institute of Pharmaceutical Science & Technology, Hanyang University)
  • 투고 : 2018.05.29
  • 심사 : 2018.08.27
  • 발행 : 2018.10.28
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초록

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

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