2AHC

Chorismate lyase with inhibitor Vanilate

2AHC の概要

• エントリーDOI: 10.2210/pdb2ahc/pdb
• 関連するPDBエントリー: 1tt8 1xlr
• 分子名称: Chorismate lyase, 4-HYDROXY-3-METHOXYBENZOATE (3 entities in total)
• 機能のキーワード: unique fold, ubiquinone pathway, 123654 antiparallel sheet, internal active site, lyase
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cytoplasm: P26602
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 75210.78

構造登録者

Gallagher, D.T.,Smith, N.N. (登録日: 2005-07-27, 公開日: 2006-01-31, 最終更新日: 2023-08-23)

主引用文献

Smith, N.N.,Roitberg, A.E.,Rivera, E.,Howard, A.,Holden, M.J.,Mayhew, M.,Kaistha, S.,Gallagher, D.T.
Structural analysis of ligand binding and catalysis in chorismate lyase
Arch.Biochem.Biophys., 445:72-80, 2006

PubMed Abstract: Chorismate lyase (CL) removes the pyruvyl group from chorismate to provide 4-hydroxybenzoate (4HB) for the ubiquinone pathway. We previously reported the crystal structure at 1.4A resolution of the Escherichia coli CL with bound 4HB product, showing that the product is bound in an internal cavity behind two flaps. To provide a more complete basis for understanding CL's unusual ligand-binding properties and mechanism of action, we now report four crystal structures of CL mutants and inhibitor complexes, together with binding and activity measurements and molecular dynamics simulations. First, an ultrahigh resolution (1.0A) crystal structure of the CL*product complex reveals details of a substrate-sized internal cavity, also behind the flaps, near the product site. Second, a 2.4A structure of CL complexed with the inhibitor vanillate shows the flaps partly opened relative to their product-bound positions. Third, a 2.0A structure of the G90A mutant with bound product reveals the basis for tighter product binding and kinetic effects of this active site mutation. Fourth, the combination of the G90A mutation with the vanillate inhibitor produces a 1.9A structure containing two inhibitor molecules, one in the product site and the other in the adjacent cavity. The two sites are connected by a short tunnel that is partly open at each end, suggesting that CL may operate via a 2-site or tunnel mechanism.

PubMed: 16343413
DOI: 10.1016/j.abb.2005.10.026

実験手法

X-RAY DIFFRACTION (2.4 Å)