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Importance of C-26 Demethylation for Homeostatic Regulation of Brassinosteroids in Seedling Shoots of Zea mays L

옥수수 유식물 신초에서 Brassinosteroids의 항상성 조절을 위반 C-26 탈메틸 반응의 중요성

  • Published : 2006.03.01

Abstract

Regulatory mechanism for endogenous levels of castasterone (CS) and its biosynthetic precursors in shoots of maize was investigated by the use of enzyme solution prepared from the plant tissue. When [$^2H_0$]- and [$^2H_6$]-CS was used as substrates, [$^2H_0$]-26-norCS and [$^2H_3$]-28-norCS were identified as products, indicating that [$^2H_0$]- and [$^2H_6$]-CS are differently metabolized into [$^2H_0$]-26-norCS and [$^2H_3$]-28-norCS by C-26 and C-28 demethylation, respectively. This suggests that both C-26 and C-28 demethylation can be involved in CS catabolism. In fact that C-28 demethylation only occurred when isotope labeled substrate was used, however, C-26 demethylation is thought be a natural reaction occurred in the maize shoots. When 6-deoxoteasterone (6-deoxoTE) was used, 6-deoxo-26-norTE and 3-dehydro-6-deoxo-26-norTE as well as 6-deoxo-3-dehydroTE and 6-deoxotyphasterol (6-deoxoTY) were identified as enzyme products. When 6-deoxoTY was added, 6-deoxo-26-norTY as well as 6-deoxo-3-dehydroTE and 6-deoxoTE was identified as products. These indicate that C-26 demethylation of 6-deoxoTE, 6-deoxo-3-dehydroTE and 6-deoxoTY as well as a reversible C-3 epimerization from 6-deoxoTE to 6-deoxoTY intermediated by 6-deoxo-3-dehydroTE are operative in the maize shoots, demonstrating that endogenous levels of biosynthetic precursors of CS are also controlled by C-26 demethylation. Therefore, it is thought that C-26 demethylation is an important and a common deactivation process which functions to maintain steady state levels of endogenous brassinosteroids in the maize shoots.

옥수수 유식물 줄기에서 중요 BRs의 함량조절 기작을 옥수수 유식물 줄기로부터 얻어진 효소원을 이용하여 조사하였다. 먼저 활성형 BR인 CS의 대사를 [$^2H_0$]-와[$^2H_6$]-CS를 기질로 사용하여 실험한 결과 [$^2H_0$]- 와 [$^2H_6$]-CS는 각각 [$^2H_0$]-26-norCS와 [$^2H_3$]-28-norCS로 전환됨을 GC-MS 분석을 통해 확인하였으며, 이러한 두 가지의 대사과정 중 C-26 탈메틸 반응에 의한 CS에서 26-norCS로의 전환만이 생체 내에서 일어나는 반응임을 확인하였다. 이와 함께 주요 생합성 전구물질인 6-deoxoTE와 6-deoxoTY에 대해서도 같은 효소원을 이용하여 C-26 탈메틸 반응에 의한 대사를 조사한 결과 6-deoxoTE는 6-deoxo-3-dehydroTE와 6-deoxoTY로, 6-deoxoTY는 6-deoxo-3-dehydroTE와 6-deoxoTE로 전환됨을 확인함과 동시에, 6-deoxoTE는 6-deoxo-26-norTE 로, 6-deoxo-3-DHT는 3-dehydro-6-deoxo-26-norTE, 6-deoxoTY는 6-deoxo-26-norTY로 전환됨을 확인하였다. 이러한 결과들은 옥수수 유식물 줄기에서 중요 BRs가 모두 C-26 탈메틸 반응이 일어날 수 있음을 나타내는 결과로서 BRs의 C-26 탈메틸 반응이 활성형 BR뿐만 아니라 그 생합성 전구물질에도 중요한 함량조절 기작임을 확인 할 수 있었다.

Keywords

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