Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
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Identification and Purification of New Brassinosteroid-Conjugates in Arabidopsis thaliana

애기장대 내 새로운 브라시노스테로이드 결합체의 분리 및 동정

  • Joo, Se-Hwan (Department of Life Science, Chung-Ang University) ;
  • Lee, Yew (Department of Biological Science Technology, Yonsei University) ;
  • Kim, Seong-Ki (Department of Life Science, Chung-Ang University)
  • 주세환 (중앙대학교 생명과학과) ;
  • 이유 (연세대학교 생명과학기술학부) ;
  • 김성기 (중앙대학교 생명과학과)
  • Received : 2011.02.07
  • Accepted : 2011.03.09
  • Published : 2011.05.30
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Abstract

Metabolism of $^3H$-castasterone in the presence of $^{32}P$-ATP was examined by an enzyme solution prepared from A. thaliana after a reversed phased HPLC, after which a polar metabolite labeled by both $^3H$ and $^{32}P$ was obtained, suggesting that $^3H$-CS is phosphorylated by $^{32}P$-ATP. To elucidate the structure of the phosphorylated CS, the same enzyme assay was carried out with non-isotopes labeled CS and ATP. In GC-MS analysis the metabolite gave a molecular ion at m/z 664 as a bismethanboronate, suggesting the metabolite is a CS phosphate. Treatment of wheat germ acid phosphatase that hydrolyzed phosphoester bond gave the same mass spectrum and GC retention time in GC-MS analyses, confirming that the metabolite is phosphorylated CS. This is the first example of phosphorylated conjugates of CS in plants.

애기장대에서 추출한 효소원에 동위원소로 표지 된 $^{32}P$-ATP와 $^3H$-castasterone를 기질로 하여 효소반응을 수행하여 얻어진 생성물을 분석한 결과 $^3H$와 $^{32}P$의 활성이 동시에 검출되었다. 이 대사산물의 구조를 알아보고자 동위원소로 표지 되지 않은 CS, ATP를 이용하여 효소 반응하여 얻어진 대사산물의 GC-MS 분석결과 ion m/z 664의 값을 얻었는데, 이를 CS phosphate로 예상할 수 있었다. 상기의 대사산물 분획에 wheat germ acid phosphatase를 처리하여 phosphoester 결합을 깨뜨린 후 얻어진 생성물의 GC-MS 분석결과 CS의 spectrum을 얻을 수 있었는데 이를 통해 식물체 내 CS의 결합체로 인산화 된 형태가 존재할 가능성을 확인하였다.

Keywords

References

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