cDNA cloning, expression and homology modeling of a luciferase from the firefly Lampyroidea maculata.
The cDNA of a firefly luciferase from lantern mRNA of Lampyroidea maculata has been cloned, sequenced and functionally expressed. The cDNA has an open reading frame of 1647 bp and codes for a 548-residue-long polypeptide. Noteworthy, sequence comparison as well as homology modeling showed the highest degree of similarity with H. unmunsana and L. mingrelica luciferases, suggesting a close phylogenetic relationship despite the geographical distance separation. The deduced amino acid sequence of the luciferase gene of firefly L. maculata showed 93% identity to H. unmunsana. Superposition of the three-dimensional model of L. maculata luciferase (generated by homology modeling) and three dimensional structure of Photinus pyralis luciferase revealed that the spatial arrangements of Luciferin and ATP-binding residues are very similar. Putative signature of AMPbinding domain among the various firefly species and Lampyroidea maculata was compared and a striking similarity was found. Different motifs and sites have been identified in Lampyroidea maculata by sequence analysis. Expression and purification of luciferase from Lampyroidea maculata was carried out using Ni-NTA Sepharose. Bioluminescence emission spectrum was similar to Photinus pyralis luciferase.
Emamzadeh
Abdo Rahman
AR
Department of Biochemistry, Faculty of Basic Sciences, Tarbiat Modarres University, Tehran, Iran.
Hosseinkhani
Saman
S
Sadeghizadeh
Majid
M
Nikkhah
Maryam
M
Chaichi
Mohammad Javad
MJ
Mortazavi
Mojtaba
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D048888:Fireflies / Q000201:enzymology* / Q000235:genetics*; D008156:Luciferases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D040681:Structural Homology, Protein
D018076:DNA, Complementary; D008156:Luciferases