한국수산과학회지 (Korean Journal of Fisheries and Aquatic Sciences)

  • 제47권4호
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  • Pages.370-375
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  • 2014
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  • 0374-8111(pISSN)
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  • 2287-8815(eISSN)
한국수산과학회 (The Korean Society of Fisheries and Aquatic Science)
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BACE2의 대량발현 및 리폴딩

Overexpression and Refolding of BACE2

  • Park, Sun Joo (Department of Chemistry, Pukyong National University) ;
  • Tai, Shuaiqi (Department of Food Science and Biotechnology, Kunsan National University) ;
  • Lee, Yeon-Ji (Department of Food Science and Biotechnology, Kunsan National University) ;
  • Jeon, You-Jin (School of Marine Biomedical Sciences, Jeju National University) ;
  • Kim, Yong-Tae (Department of Food Science and Biotechnology, Kunsan National University)
  • 투고 : 2014.07.25
  • 심사 : 2014.08.14
  • 발행 : 2014.08.31
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초록

BACE2 is a membrane-bound aspartic protease that is highly homologous with BACE1. While BACE1 processes the amyloid precursor protein (APP) at a key step in generating ${\beta}$-amyloid peptide and presumably causes Alzheimer's disease (AD), BACE2 has not been demonstrated to be involved in APP processing directly, and its physiological functions are unknown. To determine its function and to develop inhibitors from marine sources, we constructed an overexpression vector for producing BACE2. The gene encoding human BACE2 protease was amplified using the polymerase chain reaction and cloned into the pET11a expression vector, resulting in pET11a/BACE2. Recombinant BACE2 protease was overexpressed successfully in E. coli as inclusion bodies, refolded using the rapid-dilution method, and purified via two-step fast protein liquid chromatography using Sephacryl S-300 gel filtration and Resource-Q column chromatography. The BACE2 protease produced was an active form. This study provides an efficient method not only for studying the basic properties of BACE2, but also for developing inhibitors from natural marine sources.

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