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Detection of Protein S-Acylation using Acyl-Resin Assisted Capture
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ポリケチド・ディゾラゾール生物合成におけるトランス作用のアチルトランスフェラーゼとアチルキャリアタンパク質のタンパク質間相互作用の構造的基礎
Akimasa Miyanaga1, Risako Ouchi1, Fumihiro Ishikawa2
1Department of Chemistry , Tokyo Institute of Technology , 2-12-1, O-okayama , Meguro-ku, Tokyo 152-8551 , Japan.
Journal of the American Chemical Society
|June 6, 2018
まとめ
研究者はポリケチド生物合成における重要な酵素複合体の結晶構造を決定した. これは,アシルトランスフェラーゼ (AT) がアシルトランスフェラータンパク質 (ACP) を認識し,自然な製品組成の洞察を提供することを明らかにします.
科学分野:
- 生物化学
- 構造生物学
- 自然製品の生物合成
背景:
- アチルトランスフェラーゼ (AT) は,ポリケチド天然産物生物合成における重要な酵素である.
- モジュラーポリケチド合成におけるトランス作用のATは,アシル単位をアシルキャリアタンパク質 (ACP) に負荷する.
- AT-ACPの相互作用を理解することは不可欠ですが,一時的な結合のために困難です.
研究 の 目的:
- トランスアクティブのATとその同類のACPの相互作用の構造的基盤を明らかにする.
- トランスアクティブATによるACP承認のダイナミックメカニズムを特徴づける.
主な方法:
- ディゾラゾールAT-ACP複合体の結晶構造を2.03 Åの解像度で決定した.
- ブロモアセタミド・パンテイン・クロスリンク・プローブと システイン・ミューテーションを使って トランジエント・コンプレックスを安定させました
- 特定されたACP認識サイトを 変異研究で検証した.
主要な成果:
- ディソラゾールAT-ACP複合体の結晶構造は,うまく解消されました.
- ループ1とヘリックスIIIを含む特定のACP残留物は,ディゾラゾールATの主要な認識部位として特定されました.
- 変異分析は,AT-ACPの認識におけるこれらの残留物の役割を確認した.
結論:
- 決定されたAT-ACP複合構造は,トランス作用のATダイナミクスの基礎的な理解を提供します.
- トランスアクティブのATによるACP承認の詳細なメカニズムが得られた.
- この研究は,ポリケチド組立ラインのメカニズムに関するさらなる研究への道を開く.
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