4O8C

Structure of the H170Y mutant of thermostable p-nitrophenylphosphatase from Bacillus Stearothermophilus

4O8C の概要

• エントリーDOI: 10.2210/pdb4o8c/pdb
• 関連するPDBエントリー: 4KN8
• 分子名称: Thermostable NPPase, MAGNESIUM ION, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: phosphatase, hydrolase
• 由来する生物種: Geobacillus stearothermophilus
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 58398.64

構造登録者

Shen, T.,Guo, Z.,Wang, F.,Gong, W.,Ji, C. (登録日: 2013-12-27, 公開日: 2014-06-18, 最終更新日: 2023-11-08)

主引用文献

Shen, T.,Guo, Z.,Ji, C.
Structure of a His170Tyr mutant of thermostable pNPPase from Geobacillus stearothermophilus.
Acta Crystallogr.,Sect.F, 70:697-702, 2014

PubMed Abstract: Using directed evolution based on random mutagenesis and heat-treated selection, a thermostable His170Tyr mutant of Geobacillus stearothermophilus thermostable p-nitrophenylphosphatase (TpNPPase) was obtained. The temperature at which the His170Tyr mutant lost 50% of its activity (T1/2) was found to be 4.40 K higher than that of wild-type TpNPPase, and the melting temperature of the His170Tyr mutant increased by 2.39 K. The crystal structure of the His170Tyr mutant was then determined at 2.0 Å resolution in the presence of a sodium ion and a sulfate ion in the active site. The cap domain of chain B shows a half-closed conformation. The hydrophobic side chain of the mutated residue, the hydroxyphenyl group, forms a hydrophobic contact with the methyl group of Ala166. This hydrophobic interaction was found using the Protein Interactions Calculator (PIC) web server with an interaction distance of 4.6 Å, and might be a key factor in the thermostabilization of the His170Tyr mutant. This study potentially offers a molecular basis for both investigation of the catalytic mechanism and thermostable protein engineering.

PubMed: 24915075
DOI: 10.1107/S2053230X14007341

実験手法

X-RAY DIFFRACTION (2 Å)