4L0K

Crystal structure of a type II restriction endonuclease

4L0K の概要

• エントリーDOI: 10.2210/pdb4l0k/pdb
• 分子名称: DraIII (2 entities in total)
• 機能のキーワード: draiii, restriction endonuclease, reases, star activity, hydrolase
• 由来する生物種: Deinococcus radiophilus
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 103011.42

構造登録者

Zhuo, W.,Ge, J.,Yang, M. (登録日: 2013-05-31, 公開日: 2014-05-14, 最終更新日: 2024-11-20)

主引用文献

Zhuo, W.,Lai, X.,Zhang, L.,Chan, S.H.,Li, F.,Zhu, Z.,Yang, M.,Sun, D.
Elimination of inter-domain interactions increases the cleavage fidelity of the restriction endonuclease DraIII.
Protein Cell, 5:357-368, 2014

PubMed Abstract: DraIII is a type IIP restriction endonucleases (REases) that recognizes and creates a double strand break within the gapped palindromic sequence CAC↑NNN↓GTG of double-stranded DNA (↑ indicates nicking on the bottom strand; ↓ indicates nicking on the top strand). However, wild type DraIII shows significant star activity. In this study, it was found that the prominent star site is CAT↑GTT↓GTG, consisting of a star 5' half (CAT) and a canonical 3' half (GTG). DraIII nicks the 3' canonical half site at a faster rate than the 5' star half site, in contrast to the similar rate with the canonical full site. The crystal structure of the DraIII protein was solved. It indicated, as supported by mutagenesis, that DraIII possesses a ββα-metal HNH active site. The structure revealed extensive intra-molecular interactions between the N-terminal domain and the C-terminal domain containing the HNH active site. Disruptions of these interactions through site-directed mutagenesis drastically increased cleavage fidelity. The understanding of fidelity mechanisms will enable generation of high fidelity REases.

PubMed: 24733184
DOI: 10.1007/s13238-014-0038-z

実験手法

X-RAY DIFFRACTION (2.328 Å)