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In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
Published on: July 25, 2019
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ストカスティックゲートダイナミクスは,ユビキチン化酵素の触媒活性を調節する
Manoj K Rout1, Curtis D Hodge, Craig J Markin
1Department of Biochemistry, University of Alberta , Edmonton, Alberta T6G 2H7, Canada.
Journal of the American Chemical Society
|November 26, 2014
まとめ
Ubc13酵素の柔軟なループダイナミクスは,DNA損傷と免疫反応に不可欠な触媒活動を制御する. このストカスティックゲートメカニズムは,細胞シグナル伝達に不可欠なLys 63結合ポリユビキチン鎖合成を調節する.
科学分野:
- バイオケミストリー バイオケミストリー
- 分子生物学は分子生物学である.
- セルラー・シグナリング
背景:
- DNAの損傷と先天的な免疫反応は,Lys 63結合ポリユビキチン (polyUb) 鎖に依存しています.
- Ubc13は,ユビキチン結合酵素 (E2) で,その活性部位システインを介してこれらの重要なポリウブ鎖を合成します.
研究 の 目的:
- Ubc13.の触媒活性におけるアクティブサイトループダイナミクスの役割を調査する.
- ループの動きがLys 63結合型ポリユビキチン鎖の合成をどのように制御するかを理解する.
主な方法:
- Ubc13活性サイトループの残留物のサイト指向性変異.
- 酵素動力学アッセイ 酵素動力学アッセイ
- 核磁共振 (NMR) のリラックス測定.
- X線結晶学.X線結晶学.X線結晶学.
- 分子力学 (MD) シミュレーション.
- イーストにおける in vivo 機能的補完性アッセイ
主要な成果:
- アクティブサイトループのピコからナノ秒の急速な変動は,ストキャスティックゲートとして作用し,Ubc13の触媒活動を調節します.
- ループの正確なバランスの取れた開閉率は,酵素の機能にとって極めて重要です.
- ループのダイナミクスを影響する突然変異は,in vivoにおいて深刻な生物学的結果をもたらします.
結論:
- アクティブ・サイト・ループのダイナミクスは,Ubc13.の触媒的調節に極めて重要です.
- この調節メカニズムは,Lys 63結合ポリユビキチン鎖の合成に不可欠であり,DNA損傷と免疫反応に影響を与えます.
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