C-terminal region of the active domain enhances enzymatic activity in dinoflagellate luciferase.
The dinoflagellate luciferase of Lingulodinium polyedrum has three catalytic domains in its single polypeptide chain (M(r) = 137 kDa), and each 42 kDa domain is enzymatically active. Deletion mutants for N- or C-terminal regions of domain 3 of the luciferase, ranging from 29 to 38 kDa, were constructed and expressed in E. coli cells. The activities of N-terminal deleted mutants were above 20% of wild type, but showed different pH-activity profiles. By contrast, the activities of C-terminal deleted mutants decreased drastically to below 1% of wild type, although their pH-activity profiles and spectra were identical to those of wild type L. polyedrum luciferase. These results indicate that the C-terminal region of this enzyme could be important for the bioluminescence reaction, although based on crystal structure of the luciferase domain, this region does not contain active or regulatory sites.
Suzuki-Ogoh
Chie
C
Research Institute for Cell Engineering, National Institute for Advanced Industrial Science and Technology (AIST), Osaka, 563-8577, Japan.
Wu
Chun
C
Ohmiya
Yoshihiro
Y
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D017931:DNA Primers; D004141:Dinoflagellida / Q000201:enzymology*; D008156:Luciferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D017931:DNA Primers; D011994:Recombinant Proteins; D008156:Luciferases