Kuwahara_2008_Protein.Eng.Des.Sel_21_737

Reference

Title : Importance of the Ca2+-binding sites in the N-catalytic domain of a family I.3 lipase for activity and stability - Kuwahara_2008_Protein.Eng.Des.Sel_21_737
Author(s) : Kuwahara K , Angkawidjaja C , Matsumura H , Koga Y , Takano K , Kanaya S
Ref : Protein Engineering Des Sel , 21 :737 , 2008
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.

PubMedSearch : Kuwahara_2008_Protein.Eng.Des.Sel_21_737
PubMedID: 18987131
Gene_locus related to this paper: psesp-Q9RBY1

Related information

Gene_locus psesp-Q9RBY1
Family psesp-Q9RBY1    Bacterial_lipase    Bacterial_lip_FamI.3
Structure psesp-Q9RBY1    Bacterial_lipase    Bacterial_lip_FamI.3    2ZJ6    2ZJ7

Citations formats

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

Kuwahara K, Angkawidjaja C, Matsumura H, Koga Y, Takano K, Kanaya S (2008)
Protein Engineering Des Sel 21 :737