3SKD

Crystal structure of the Thermus thermophilus cas3 HD domain in the presence of Ni2+

3SKD の概要

• エントリーDOI: 10.2210/pdb3skd/pdb
• 分子名称: Putative uncharacterized protein TTHB187, NICKEL (II) ION (3 entities in total)
• 機能のキーワード: crispr, cas, hd domain, nuclease, hydrolase
• 由来する生物種: Thermus thermophilus HB8
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29781.69

構造登録者

Bailey, S.,Mulepati, S. (登録日: 2011-06-22, 公開日: 2011-07-20, 最終更新日: 2024-02-28)

主引用文献

Mulepati, S.,Bailey, S.
Structural and Biochemical Analysis of Nuclease Domain of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated Protein 3 (Cas3).
J.Biol.Chem., 286:31896-31903, 2011

PubMed Abstract: RNA transcribed from clustered regularly interspaced short palindromic repeats (CRISPRs) protects many prokaryotes from invasion by foreign DNA such as viruses, conjugative plasmids, and transposable elements. Cas3 (CRISPR-associated protein 3) is essential for this CRISPR protection and is thought to mediate cleavage of the foreign DNA through its N-terminal histidine-aspartate (HD) domain. We report here the 1.8 Å crystal structure of the HD domain of Cas3 from Thermus thermophilus HB8. Structural and biochemical studies predict that this enzyme binds two metal ions at its active site. We also demonstrate that the single-stranded DNA endonuclease activity of this T. thermophilus domain is activated not by magnesium but by transition metal ions such as manganese and nickel. Structure-guided mutagenesis confirms the importance of the metal-binding residues for the nuclease activity and identifies other active site residues. Overall, these results provide a framework for understanding the role of Cas3 in the CRISPR system.

PubMed: 21775431
DOI: 10.1074/jbc.M111.270017

実験手法

X-RAY DIFFRACTION (2 Å)