Importance of a repetitive nine-residue sequence motif for intracellular stability and functional structure of a family I.3 lipase.
PML5 is a functional derivative of a family I.3 lipase from Pseudomonas sp. MIS38 and contains five repeats of a nine-residue sequence motif. Two aspartate residues within the second and third repetitive sequences of PML5 were replaced by Ala. The secretion level, intracellular accumulation level, and stability of the resultant mutant protein were greatly reduced as compared to those of PML5. In addition, this mutant protein was inactive and did not bind Ca2+ ion. We propose that the repetitive sequences of PML5 form a beta-roll structure in the cells and thereby contribute to the intracellular stability and secretion efficiency of the protein.
Angkawidjaja
Clement
C
Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Paul
Aditya
A
Koga
Yuichi
Y
Takano
Kazufumi
K
Kanaya
Shigenori
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002151:Calorimetry; D002942:Circular Dichroism; D004795:Enzyme Stability; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D011549:Pseudomonas / Q000201:enzymology
D001426:Bacterial Proteins; D008049:Lipase; D002118:Calcium