Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Construction of a Transgenic Plant to Develop a New Method for the Isolation of Calmodulin-Binding Proteins

새로운 방법을 이용한 칼모둘린 결합 단백질 분리를 위한 형질 전환 식물체의 구축

  • Kim, Sun-Ho (Environmental Biotechnology National Core Research Center, Gyeongsang National University) ;
  • Lee, Kyung-Hee (Environmental Biotechnology National Core Research Center, Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Kim, Kyung-Eun (Division of Applied Life Science (BK21 program), Plant Molecular Bilogy and Biotechnology Research Center, Gyeongsang National University) ;
  • Jung, Mi-Soon (Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Lim, Chae-Oh (Environmental Biotechnology National Core Research Center, Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Lee, Shin-Woo (Department of Crops Biotechnology, Jinju National University) ;
  • Chung, Woo-Sik (Environmental Biotechnology National Core Research Center, Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University)
  • 김선호 (경상대학교 환경생명과학 국가핵심연구센터) ;
  • 이경희 (경상대학교 환경생명과학 국가핵심연구센터, 응용생명과학부) ;
  • 김경은 (경상대학교 응용생명과학부) ;
  • 정미순 (경상대학교 응용생명과학부) ;
  • 임채오 (경상대학교 환경생명과학 국가핵심연구센터, 응용생명과학부) ;
  • 이신우 (진주산업대학교 작물과학과) ;
  • 정우식 (경상대학교 환경생명과학 국가핵심연구센터, 응용생명과학부)
  • Published : 2007.09.30
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Abstract

Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety CaM-binding proteins (CaMBPs). Because eukaryotes have multiple CaMBPs, it is important to isolate and characterize them in different tissues and conditions. So far a number of CaMBPs have been identified through classical screening methods. Many classes of proteins have been predicted to bind CaMs based on their structural homology with already known targets. In an effort to develop a method for large-scale analysis of CaMBPs in Arabidopsis, we have generated a transgenic plants overexpressing AtCaM2-GFP. We performed protein pull-down assay to test whether exogenously expressed AtCaM2-GFP proteins can interact with CaMBPs. The exogenously expressed AtCaM2-GFP could strongly interact with a CaMBP, AS1 protein. This result suggests that AtCaM2-GFP in transgenic plants may interact with many CaMBPs in plant cell. Therefore, we will be able to isolate kinds of CaMBPs by using these transgenic plants in many different tissue and environments.

칼모둘린은 칼슘과 결합하는 센서로써 다양한 칼모둘린 결합 단백질들과의 상호 작용을 통하여 세포 내에서 여러가지 기능을 조절한다. 진핵 생물들은 많은 종류의 칼모둘린 결합 단백질을 가지고 있기 때문에 이러한 단백질들의 분리와 특성 규명이 중요하다. 이미 여러 가지 방법들을 이용하여 칼모둘린 결합 단백질들이 분리되었고 이미 알려진 단백질의 구조적인 유사성을 토대로 더 많은 단백질들이 예측되었다. 우리는 애기장대에서 칼모둘린 결합 단백질의 분리와 특성 규명을 위해 형광 단백질과 융합된 칼모둘린 과발현 형질 전환체를 제조하여 공촛점 현미경과 Western blot 을 이용하여 과발현 형질 전환체를 선별하였다. 또한 형질 전환체 내의 칼모둘린이 칼모둘린 결합 단백질과 상호 작용함을 pull-down 분석을 통해서 확인하였다. 이러한 결과들을 토대로 칼모둘린 과발현 형질 전환체를 이용하여, 칼모둘린과 상호 작용하는 여러 가지 칼모둘린 결합 단백질들을 분리할 수 있을 것으로 기대된다.

Keywords

References

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