Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Bombyx mori Protein Disulfide Isomerase (bPDI) Protects Sf9 Cells from Endoplasmic Reticulum (ER) Stress

소포체 스트레스에 대한 Protein Disulfide Isomerase의 세포보호효과

  • Goo, Tae-Won (Department of Agricultural biology, the National Institute of Agriculture Science and Technology) ;
  • Yun, Eun-Young (Department of Agricultural biology, the National Institute of Agriculture Science and Technology) ;
  • Kim, Sung-Wan (Department of Agricultural biology, the National Institute of Agriculture Science and Technology) ;
  • Choi, Kwang-Ho (Department of Agricultural biology, the National Institute of Agriculture Science and Technology) ;
  • Kang, Seok-Woo (Department of Agricultural biology, the National Institute of Agriculture Science and Technology) ;
  • Kwon, Ki-Sang (Department of Anatomy, College of Medicine, Chungnam National University) ;
  • Kwon, O-Yu (Department of Anatomy, College of Medicine, Chungnam National University)
  • 구태원 (농업과학기술원 농업생물부) ;
  • 윤은영 (농업과학기술원 농업생물부) ;
  • 김성완 (농업과학기술원 농업생물부) ;
  • 최광호 (농업과학기술원 농업생물부) ;
  • 강석우 (농업과학기술원 농업생물부) ;
  • 권기상 (충남대학교 의과대학 해부학교실) ;
  • 권오유 (충남대학교 의과대학 해부학교실)
  • Published : 2007.08.30
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Abstract

In the previous our study, a cDNA that encodes protein disulfide isomerase from Bombyx mori (bPDI)was isolated and characterized. bPDI has an open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and ER (endoplasmic reticulum) retention signal of the KDEL motif at its C-terminal. Recent studies have demonstrated that misfolded proteins are accumulated in many diseases including Alzheimer’s, goiter, emphysema, and prion infections. bPDI was over-expressed or knock-downed in Sf9 cells to study the relationship between bPDI expression and protections against protein misfolding. bPDI gene was cloned in insect expression vector pIZT/V5-His for over-expression and bPDI double-stranded RNA (dsRNA) was generated for knock-down. Over-expression of bPDI significantly improved survival rate, but bPDI dsRNA transfection significantly reduced survival rate after 48 hours exposure. In mock-transfected or wild-type cells had no significant effect. The results support the view that bPDI is one of the important intracellular components for cell protect mechanism, especially, against ER stress such as protein misfolding.

bPDI가 ER내 misfolding 단백질의 생성을 제한함으로써 곤충변역과 관계하는지를 해석하기 위하여 bPDI가 과발현(overexpression)되는 곤충세포주와 이와 반대로 bPDI가 억제발현(knock-down)되는 곤충세포주를 제작하여 bPDI가 곤충면역에 관련하는지를 해석하였다. bPDI가 과발현되는 세포주 (Sf9-bPDI)는 정상세포주(Sf9)나 pIZT/V5-His 벡터만 도입된 세포주(Sf9-pIZT)에 비하여 생존율이 30% 이상 높았지만, bPDI의 전사체 발현이 억제된 세포주(Sf9-bPDI-dsRNA)는 오히려 정상세포주나 pIZT/V5-His 벡터만 도입된 세포주에 비하여 생존율이 약 15%낮았다. 이와 같은 결과로써, bPDI는 ER내 misfolding 단백질의 생성을 제한함으로써 곤충의 ERSE과 밀접하게 관련할 것이라 추정할 수 있었다.

Keywords

References

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