3FWH

Structure of haloalkane dehalogenase mutant Dha15 (I135F/C176Y) from Rhodococcus rhodochrous

3FWH の概要

• エントリーDOI: 10.2210/pdb3fwh/pdb
• 関連するPDBエントリー: 3FBW 3G9X
• 分子名称: Haloalkane dehalogenase, ACETATE ION, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: alpha/beta hydrolase core, helical cap domain, catalytic triad (asp106, his272, glu130), mutant, i135f, c176y, haloalkanes, detoxification, hydrolase
• 由来する生物種: Rhodococcus sp.
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34394.96

構造登録者

Gavira, J.A.,Stsiapanava, A.,Kuty, M.,Dohnalek, J.,Lapkouski, M.,Kuta Smatanova, I. (登録日: 2009-01-18, 公開日: 2010-02-02, 最終更新日: 2023-09-06)

主引用文献

Stsiapanava, A.,Dohnalek, J.,Gavira, J.A.,Kuty, M.,Koudelakova, T.,Damborsky, J.,Kuta Smatanova, I.
Atomic resolution studies of haloalkane dehalogenases DhaA04, DhaA14 and DhaA15 with engineered access tunnels.
Acta Crystallogr.,Sect.D, 66:962-969, 2010

PubMed Abstract: The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 is a bacterial enzyme that shows catalytic activity for the hydrolytic degradation of the highly toxic industrial pollutant 1,2,3-trichloropropane (TCP). Mutagenesis focused on the access tunnels of DhaA produced protein variants with significantly improved activity towards TCP. Three mutants of DhaA named DhaA04 (C176Y), DhaA14 (I135F) and DhaA15 (C176Y + I135F) were constructed in order to study the functional relevance of the tunnels connecting the buried active site of the protein with the surrounding solvent. All three protein variants were crystallized using the sitting-drop vapour-diffusion technique. The crystals of DhaA04 belonged to the orthorhombic space group P2(1)2(1)2(1), while the crystals of DhaA14 and DhaA15 had triclinic symmetry in space group P1. The crystal structures of DhaA04, DhaA14 and DhaA15 with ligands present in the active site were solved and refined using diffraction data to 1.23, 0.95 and 1.22 A, resolution, respectively. Structural comparisons of the wild type and the three mutants suggest that the tunnels play a key role in the processes of ligand exchange between the buried active site and the surrounding solvent.

PubMed: 20823547
DOI: 10.1107/S0907444910027101

実験手法

X-RAY DIFFRACTION (1.22 Å)