1P6C

crystal structure of phosphotriesterase triple mutant H254G/H257W/L303T complexed with diisopropylmethylphosphonate

1P6C の概要

• エントリーDOI: 10.2210/pdb1p6c/pdb
• 関連するPDBエントリー: 1P6B
• 分子名称: Parathion hydrolase, ZINC ION, DIETHYL 4-METHYLBENZYLPHOSPHONATE, ... (5 entities in total)
• 機能のキーワード: metalloenzyme, tim barrel, nerve agent, hydrolase
• 由来する生物種: Flavobacterium sp.
• 細胞内の位置: Cell membrane ; Peripheral membrane protein : P0A433
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 73767.13

構造登録者

Hill, C.M.,Li, W.,Thoden, J.B.,Holden, H.M.,Raushel, F.M. (登録日: 2003-04-29, 公開日: 2003-09-16, 最終更新日: 2023-11-15)

主引用文献

Hill, C.M.,Li, W.S.,Thoden, J.B.,Holden, H.M.,Raushel, F.M.
Enhanced degradation of chemical warfare agents through molecular engineering of the phosphotriesterase active site.
J.Am.Chem.Soc., 125:8990-8991, 2003

PubMed Abstract: The bacterial phosphotriesterase has been utilized as a template for the evolution of improved enzymes for the catalytic decomposition of organophosphate nerve agents. A combinatorial library of active site mutants was constructed by randomizing residues His-254, His-257, and Leu-303. The collection of mutant proteins was screened for the ability to hydrolyze a chromogenic analogue of the most toxic stereoisomer of the chemical warfare agent, soman. The mutant H254G/H257W/L303T catalyzed the hydrolysis of the target substrate nearly 3 orders of magnitude faster than the wild-type enzyme. The X-ray crystal structure was solved in the presence and absence of diisopropyl methyl phosphonate. The mutant enzyme was ligated to an additional divalent cation at the active site that was displaced upon the binding of the substrate analogue inhibitor. These studies demonstrate that substantial changes in substrate specificity can be achieved by relatively minor changes to the primary amino acid sequence.

PubMed: 15369336
DOI: 10.1021/ja0358798

実験手法

X-RAY DIFFRACTION (2 Å)