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8XK3

Structure of the Argonaute protein from Kurthia massiliensis in complex with guide DNA and 19-mer target DNA in target-cleaved state

8XK3 の概要
エントリーDOI10.2210/pdb8xk3/pdb
EMDBエントリー38414
分子名称KmAgo, guide DNA, target DNA-1, ... (6 entities in total)
機能のキーワードmesophilic prokaryotic argonaute, dna binding protein
由来する生物種Kurthia massiliensis
詳細
タンパク質・核酸の鎖数4
化学式量合計96939.45
構造登録者
Tao, X.,Ding, H.,Wu, S. (登録日: 2023-12-22, 公開日: 2025-01-01, 最終更新日: 2025-07-16)
主引用文献Tao, X.,Ding, H.,Wu, S.,Wang, F.,Xu, H.,Li, J.,Zhai, C.,Li, S.,Chen, K.,Wu, S.,Liu, Y.,Ma, L.
Structural and mechanistic insights into a mesophilic prokaryotic Argonaute.
Nucleic Acids Res., 52:11895-11910, 2024
Cited by
PubMed Abstract: Argonaute (Ago) proteins are programmable nucleases found in all domains of life, playing a crucial role in biological processes like DNA/RNA interference and gene regulation. Mesophilic prokaryotic Agos (pAgos) have gained increasing research interest due to their broad range of potential applications, yet their molecular mechanisms remain poorly understood. Here, we present seven cryo-electron microscopy structures of Kurthia massiliensis Ago (KmAgo) in various states. These structures encompass the steps of apo-form, guide binding, target recognition, cleavage, and release, revealing that KmAgo employs a unique DDD catalytic triad, instead of a DEDD tetrad, for DNA target cleavage under 5'P-DNA guide conditions. Notably, the last catalytic residue, D713, is positioned outside the catalytic pocket in the absence of guide. After guide binding, D713 enters the catalytic pocket. In contrast, the corresponding catalytic residue in other Agos has been consistently located in the catalytic pocket. Moreover, we identified several sites exhibiting enhanced catalytic activity through alanine mutagenesis. These sites have the potential to serve as engineering targets for augmenting the catalytic efficiency of KmAgo. This structural analysis of KmAgo advances the understanding of the diversity of molecular mechanisms by Agos, offering insights for developing and optimizing mesophilic pAgos-based programmable DNA and RNA manipulation tools.
PubMed: 39315697
DOI: 10.1093/nar/gkae820
主引用文献が同じPDBエントリー
実験手法
ELECTRON MICROSCOPY (2.76 Å)
構造検証レポート
Validation report summary of 8xk3
検証レポート(詳細版)ダウンロードをダウンロード

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件を2025-07-23に公開中

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