配合的に結合されたSUMO-E2-E3リガースのテルナー複合体の形成のための原子的に調整されたSUMO E2結合酵素の化学合成

Yinfeng Zhang¹, Tsuyoshi Hirota¹, Keiko Kuwata¹

¹Institute of Transformative Bio-Molecules (WPI-ITbM) , Nagoya University , Chikusa , Nagoya 464-8602 , Japan.

Journal of the American Chemical Society

|August 23, 2019

まとめ

研究者はSUMOylationを研究するために改造されたUbc9酵素を作成しました. このアプローチはE3リガスを成功裏に捕まえて,タンパク質結合経路の研究を可能にしました.

科学分野:

生物化学
分子生物学
化学生物学

背景:

E2結合酵素は,ユビキチンとSUMO変形剤を基板タンパク質に転送することを促進する.
高い伝達率とE2酵素の不規則な結合は,それらの相互作用の研究を妨げます.
E2酵素結合パートナーを調査するには,特定の安定した結合体が必要です.

研究の目的:

SUMO E2 結合酵素の相互作用を研究するために,合成 Ubc9 変種を開発する.
特定の E2-SUMO 結合体にアクセスするためのツールを作成します.
Ubc9と相互作用するE3リガスを捕まえて識別する.

主な方法:

安定したアミド結合,光反応性グループ,およびバイオチンタグを持つ Ubc9のサイト特異的改変.
結合されたネイティブ化学結合 (NCL),α-ケト酸-ヒドロキシラミン結合 (KAHA),およびセリン/スレオニン結合 (STL).
合成タンパク質化学を用いて Ubc9の変異体を作りました

主要な成果:

望ましい修正を伴う Ubc9 変種を成功裏に作成した.
Ubc9-SUMO結合体の選択的形成が示されている.
安定したSUMO1-Ubc9-RanBP2三重複合体を形成するE3リガース (RanBP2) を捕らえた.

結論:

結合方法の強力な組み合わせは,機能化されたE2結合酵素の合成を可能にします.
このアプローチは,E3リガスを捕まえるための化学的探査機の開発を容易にする.
この方法論は,他のタンパク質を合成し,それらの相互作用を研究するために適用できます.

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