1F0O

PVUII ENDONUCLEASE/COGNATE DNA COMPLEX (GLUTARALDEHYDE-CROSSLINKED CRYSTAL) AT PH 7.5 WITH TWO CALCIUM IONS AT EACH ACTIVE SITE

1F0O の概要

• エントリーDOI: 10.2210/pdb1f0o/pdb
• 関連するPDBエントリー: 1EYU 1PVI 2PYI 3PYI
• 分子名称: (5'-D(*TP*GP*AP*CP*CP*AP*GP*CP*TP*GP*GP*TP*C-3'), TYPE II RESTRICTION ENZYME PVUII, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: protein-dna complex, endonuclease type ii, restriction enzyme, catalytic metal visualization, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Proteus vulgaris
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 44837.47

構造登録者

Horton, J.R.,Cheng, X. (登録日: 2000-05-16, 公開日: 2000-11-06, 最終更新日: 2024-02-07)

主引用文献

Horton, J.R.,Cheng, X.
PvuII endonuclease contains two calcium ions in active sites.
J.Mol.Biol., 300:1049-1056, 2000

PubMed Abstract: Restriction endonucleases differ in their use of metal cofactors despite having remarkably similar folds for their catalytic regions. To explore this, we have characterized the interaction of endonuclease PvuII with the catalytically incompetent cation Ca(2+). The structure of a glutaraldehyde-crosslinked crystal of the endonuclease PvuII-DNA complex, determined in the presence of Ca(2+) at a pH of approximately 6.5, supports a two-metal mechanism of DNA cleavage by PvuII. The first Ca(2+) position matches that found in all structurally examined endonucleases, while the second position is similar to that of EcoRV but is distinct from that of BamHI and BglI. The location of the second metal in PvuII, unlike that in BamHI/BglI, permits no direct interaction between the second metal and the O3' oxygen leaving group. However, the interactions between the DNA scissile phosphate and the metals, the first metal and the attacking water, and the attacking water and DNA are the same in PvuII as they are in the two-metal models of BamHI and BglI, but are distinct from the proposed three-metal or the two-metal models of EcoRV.

PubMed: 10903853
DOI: 10.1006/jmbi.2000.3938

実験手法

X-RAY DIFFRACTION (2.5 Å)