2ZJ7

Crystal structure of D157A mutant of Pseudomonas sp. MIS38 lipase

2ZJ7 の概要

• エントリーDOI: 10.2210/pdb2zj7/pdb
• 関連するPDBエントリー: 2Z8X 2Z8Z 2ZJ6
• 分子名称: Lipase, CALCIUM ION, ZINC ION, ... (4 entities in total)
• 機能のキーワード: family i.3 lipase, beta roll, calcium binding protein, rtx protein, hydrolase, calcium site mutants
• 由来する生物種: Pseudomonas sp.
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 65051.45

構造登録者

Angkawidjaja, C.,Kuwahara, K.,Kanaya, S. (登録日: 2008-02-29, 公開日: 2008-12-02, 最終更新日: 2023-11-01)

主引用文献

Kuwahara, K.,Angkawidjaja, C.,Matsumura, H.,Koga, Y.,Takano, K.,Kanaya, S.
Importance of the Ca2+-binding sites in the N-catalytic domain of a family I.3 lipase for activity and stability
Protein Eng.Des.Sel., 21:737-744, 2008

PubMed Abstract: A family I.3 lipase from Pseudomonas sp. MIS38 (PML) contains three Ca(2+)-binding sites (Ca1-Ca3) in the N-catalytic domain. Of them, the Ca1 site is formed only in an open conformation. To analyze the role of these Ca(2+)-binding sites, three mutant proteins D157A-PML, D275A-PML and D337A-PML, which are designed to remove the Ca1, Ca2 and Ca3 sites, respectively, were constructed. Of them, the crystal structures of D157A-PML and D337A-PML in a closed conformation were determined. Both structures are nearly identical to that of the wild-type protein, except that the Ca3 site is missing in the D337A-PML structure. D157A-PML was as stable as the wild-type protein. Nevertheless, it exhibited little lipase and very weak esterase activities. D275A-PML was less stable than the wild-type protein by approximately 5 degrees C in T(1/2). It exhibited weak but significant lipase and esterase activities when compared with the wild-type protein. D337A-PML was also less stable than the wild-type protein by approximately 5 degrees C in T(1/2) but was fully active. These results suggest that the Ca1 site is required to make the active site fully open by anchoring lid 1. The Ca2 and Ca3 sites contribute to the stabilization of PML. The Ca2 site is also required to make PML fully active.

PubMed: 18987131
DOI: 10.1093/protein/gzn057

実験手法

X-RAY DIFFRACTION (2.21 Å)