4ACS

Crystal structure of mutant GST A2-2 with enhanced catalytic efficiency with azathioprine

4ACS の概要

• エントリーDOI: 10.2210/pdb4acs/pdb
• 関連するPDBエントリー: 1AGS 2VCT 2WJU
• 分子名称: GLUTATHIONE S-TRANSFERASE A2, GLUTATHIONE (3 entities in total)
• 機能のキーワード: transferase, oxidation-reduction
• 由来する生物種: HOMO SAPIENS (HUMAN)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 103740.24

構造登録者

Zhang, W.,Moden, O.,Tars, K.,Mannervik, B. (登録日: 2011-12-19, 公開日: 2011-12-28, 最終更新日: 2023-12-20)

主引用文献

Zhang, W.,Moden, O.,Tars, K.,Mannervik, B.
Structure-based redesign of GST A2-2 for enhanced catalytic efficiency with azathioprine.
Chem.Biol., 19:414-421, 2012

PubMed Abstract: Glutathione transferase (GST) A2-2 is the most efficient human enzyme in the biotransformation of the prodrug azathioprine (Aza). The activation of Aza has therapeutic potential for possible use of GSTs in targeted enzyme-prodrug treatment of diseases. Based on the assumed catalytic mechanism and computational docking of Aza to the active site of the enzyme, active-site residues were selected for construction of focused mutant libraries, which were thereafter screened for Aza activity. Mutants with elevated Aza activity were identified, DNA sequenced, and the proteins purified. The two most active mutants showed up to 70-fold higher catalytic efficiency than the parental GST A2-2. The structure of the most active triple mutant (L107G/L108D/F222H) enzyme was determined by X-ray crystallography demonstrating significant changes in the topography of the active site facilitating productive binding of Aza as a substrate.

PubMed: 22444596
DOI: 10.1016/j.chembiol.2012.01.021

実験手法

X-RAY DIFFRACTION (2.1 Å)