1OBJ

Crystal structure of the T150A mutant of Malonamidase E2 from Bradyrhizobium japonicum

1OBJ の概要

• エントリーDOI: 10.2210/pdb1obj/pdb
• 関連するPDBエントリー: 1GR8 1GR9 1GRK 1O9N 1O9O 1O9P 1O9Q 1OBI 1OBK 1OBL
• 分子名称: MALONAMIDASE E2 (2 entities in total)
• 機能のキーワード: amidase
• 由来する生物種: BRADYRHIZOBIUM JAPONICUM
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 87411.45

構造登録者

Shin, S.,Oh, B.-H. (登録日: 2003-01-31, 公開日: 2003-12-18, 最終更新日: 2023-12-13)

主引用文献

Shin, S.,Yun, Y.S.,Koo, H.M.,Kim, Y.S.,Choi, K.Y.,Oh, B.-H.
Characterization of a Novel Ser-Cisser-Lys Catalytic Triad in Comparison with the Classical Ser-His-Asp Triad
J.Biol.Chem., 278:24937-, 2003

PubMed Abstract: Amidase signature family enzymes, which are widespread in nature, contain a newly identified Ser-cisSer-Lys catalytic triad in which the peptide bond between Ser131 and the preceding residue Gly130 is in a cis configuration. In order to characterize the property of the novel triad, we have determined the structures of five mutant malonamidase E2 enzymes that contain a Cys-cisSer-Lys, Ser-cisAla-Lys, or Ser-cisSer-Ala triad or a substitution of Gly130 with alanine. Cysteine cannot replace the role of Ser155 due to a hyper-reactivity of the residue, which results in the modification of the cysteine to cysteinyl sulfinic acid, most likely inside the expression host cells. The lysine residue plays a structural as well as a catalytic role, since the substitution of the residue with alanine disrupts the active site structure completely. The two observations are in sharp contrast with the consequences of the corresponding substitutions in the classical Ser-His-Asp triad. Structural data on the mutant containing the Ser-cisAla-Lys triad convincingly suggest that Ser131 plays an analogous catalytic role as the histidine of the Ser-His-Asp triad. The unusual cis configuration of Ser131 appears essential for the precise contacts of this residue with the other triad residues, as indicated by the near invariance of the preceding glycine residue (Gly130), structural data on the G130A mutant, and by a modeling experiment. The data provide a deep understanding of the role of each residue of the new triad at the atomic level and demonstrate that the new triad is a catalytic device distinctively different from the classical triad or its variants.

PubMed: 12711609
DOI: 10.1074/JBC.M302156200

実験手法

X-RAY DIFFRACTION (1.9 Å)