생명과학회지 (Journal of Life Science)

  • 제21권5호
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  • Pages.671-677
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  • 2011
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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칼모듈린에 결합하는 대두 Ca2+-ATPase 2 (SCA2)의 분리 및 특성 분석

Isolation and Characterization of a Calmodulin-binding Ca2+-ATPase 2 (SCA2) in Soybean

  • 박형철 (경상대학교 대학원 응용생명과학부, 식물분자생물학 및 유전자조작 연구소) ;
  • 김호수 (경상대학교 대학원 응용생명과학부, 식물분자생물학 및 유전자조작 연구소) ;
  • 이상민 (경상대학교 대학원 응용생명과학부, 식물분자생물학 및 유전자조작 연구소) ;
  • 조현설 (광양보건대학 물리치료과) ;
  • 정우식 (경상대학교 대학원 응용생명과학부, 식물분자생물학 및 유전자조작 연구소)
  • Park, Hyeong-Cheol (Division of Applied Life Science (BK21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Kim, Ho-Soo (Division of Applied Life Science (BK21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Lee, Sang-Min (Division of Applied Life Science (BK21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University) ;
  • Cho, Hyeon-Seol (Department of Physical Therapy, Gwangyang Health College) ;
  • Chung, Woo-Sik (Division of Applied Life Science (BK21 Program), and Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University)
  • 투고 : 2011.04.15
  • 심사 : 2011.05.13
  • 발행 : 2011.05.30
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초록

대두의 세포막에 존재하는 SCA1은 칼모듈린에 의해서 조절된다는 내용을 이전에 보고하였다. 본 연구에서는 대두의 $Ca^{2+}$-ATPase인 SCA2에 관한 특성을 연구하였다. SCA2는 SCA1과 아미노산 서열 비교에서 78%로 높은 유사성을 나타내며, 10개의 transmembrane 도메인이 존재하는 것을 확인하였다. CaM overaly assay로부터, SCA2는 칼슘에 의존적인 방법으로 칼모듈린과 결합한다는 것을 보여주었으며, Southern blot 분석 결과, 대두의 genome에는 두 종류의 $Ca^{2+}$-ATPase가 존재하는 것으로 보인다. SCA2의 $Ca^{2+}$-ATPase 효소활성을 확인하고자 yeast mutant를 이용하여 complementation assay를 수행해 보면, SCA2가 $Ca^{2+}$-ATPase의 효소활성을 가지는 것을 보여 주었다. 이러한 결과들은 SCA2가 식물에 존재하는 type IIB $Ca^{2+}$-ATPase들과 구조적으로 높은 유사성을 가진다는 것을 시사한다.

We previously reported the isolation and characterization of a gene, SCA1 (for soybean $Ca^{2+}$-ATPase 1), encoding a calmodulin-regulated $Ca^{2+}$-ATPase that is located in the plasma membrane in soybean. Here, a $Ca^{2+}$-ATPase designated as SCA2 was isolated from soybean. The two $Ca^{2+}$-ATPases, SCA1 and SCA2, share a remarkably high degree of similarity (78%). Ten transmemebrane domains were predicted by hydropathy analysis. Using gel overlay assays, CaM was found to bind to SCA2 in a $Ca^{2+}$-dependent manner. Southern blot analysis revealed the presence of two copies of the $Ca^{2+}$-ATPase gene in the soybean genome. An N-terminal truncation mutant that deletes sequence through the putative calmodulin binding site was able to complement a yeast mutant (K616) that was deficient in two endogenous $Ca^{2+}$ pumps. Our results indicate that SCA2 is structurally highly conserved with type IIB $Ca^{2+}$ pumps in plants.

키워드

참고문헌

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