1NA6

Crystal structure of restriction endonuclease EcoRII mutant R88A

1NA6 の概要

• エントリーDOI: 10.2210/pdb1na6/pdb
• 分子名称: Restriction endonuclease EcoRII (2 entities in total)
• 機能のキーワード: site-specific restriction; mutation; replication, hydrolase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 90914.98

構造登録者

Zhou, X.E.,Wang, Y.,Reuter, M.,Mucke, M.,Kruger, D.H.,Meehan, E.J.,Chen, L. (登録日: 2002-11-26, 公開日: 2003-12-16, 最終更新日: 2024-02-14)

主引用文献

Zhou, X.E.,Wang, Y.,Reuter, M.,Mucke, M.,Kruger, D.H.,Meehan, E.J.,Chen, L.
Crystal structure of type IIE restriction endonuclease EcoRII reveals an autoinhibition mechanism by a novel effector-binding fold.
J.Mol.Biol., 335:307-319, 2004

PubMed Abstract: EcoRII is a type IIE restriction endonuclease that interacts with two copies of the DNA recognition sequence 5'CCWGG, one being the actual target of cleavage, the other serving as the allosteric effector. The mode of enzyme activation by effector binding is unknown. To investigate the molecular basis of activation and cleavage mechanisms by EcoRII, the crystal structure of EcoRII mutant R88A has been solved at 2.1A resolution. The EcoRII monomer has two domains linked through a hinge loop. The N-terminal effector-binding domain has a novel DNA recognition fold with a prominent cleft. The C-terminal catalytic domain has a restriction endonuclease-like fold. Structure-based sequence alignment identified the putative catalytic site of EcoRII that is spatially blocked by the N-terminal domain. The structure together with the earlier characterized EcoRII enzyme activity enhancement in the absence of its N-terminal domain reveal an autoinhibition/activation mechanism of enzyme activity mediated by a novel effector-binding fold. This is the first case of autoinhibition, a mechanism described for many transcription factors and signal transducing proteins, of a restriction endonuclease.

PubMed: 14659759
DOI: 10.1016/j.jmb.2003.10.030

実験手法

X-RAY DIFFRACTION (2.1 Å)