| Title : Importance of an extreme C-terminal motif of a family I.3 lipase for stability - Kuwahara_2011_Protein.Eng.Des.Sel_24_411 |
| Author(s) : Kuwahara K , Angkawidjaja C , Koga Y , Takano K , Kanaya S |
| Ref : Protein Engineering Des Sel , 24 :411 , 2011 |
| Abstract : A five-residue sequence motif (VTLVG) located at positions 15-19 from the C-terminus of family I.3 lipase from Pseudomonas sp. MIS38 (PML) and an extreme C-terminal motif (DGIVIA) located at the C-terminus of PML are relatively well conserved in the passenger proteins of type 1 secretion system (T1SS). To analyze the role of these motifs, four mutant proteins of PML (PMLDelta5, PMLDelta10, 3A-PML and 2A-PML) were constructed. PMLDelta5 and PMLDelta10 lack the C-terminal 5 and 10 residues of PML, respectively. 3A-PML has triple mutations within an extreme C-terminal motif and 2A-PML has double mutations within a five-residue sequence motif. Secretion of these proteins was analyzed using Escherichia coli DH5 cells carrying Lip system (T1SS for family I.3 lipase). The secretion level of 2A-PML was dramatically reduced when compared with that of PML, whereas the secretion level of 3A-PML was comparable to that of PML, indicating that a five-residue sequence motif, instead of an extreme C-terminal motif, is required for secretion of PML. None of the mutations and truncations seriously affects the enzymatic activity of PML. However, 3A-PML, PMLDelta5 and PMLDelta10 were less stable than PML by 2.1, 7.6 and 7.6 degrees C in T(1/2), respectively, and by 5.0, 21.3 and 17.9 kJ/mol in DeltaG(H(2)O), respectively. These results indicate that an extreme C-terminal motif of PML is important for stability. |
| ESTHER : Kuwahara_2011_Protein.Eng.Des.Sel_24_411 |
| PubMedSearch : Kuwahara_2011_Protein.Eng.Des.Sel_24_411 |
| PubMedID: 21216728 |
Kuwahara K, Angkawidjaja C, Koga Y, Takano K, Kanaya S (2011)
Importance of an extreme C-terminal motif of a family I.3 lipase for stability
Protein Engineering Des Sel
24 :411
Kuwahara K, Angkawidjaja C, Koga Y, Takano K, Kanaya S (2011)
Protein Engineering Des Sel
24 :411