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Cloning of the Orange Light-Producing Luciferase from Photinus scintillans-A New Proposal on how Bioluminescence Color is Determined
- Source :
- Photochemistry and photobiology. 93(2)
- Publication Year :
- 2016
-
Abstract
- Unlike the enchanting yellow-green flashes of light produced on warm summer evenings by Photinus pyralis, the most common firefly species in North America, the orange lights of Photinus scintillans are infrequently observed. These Photinus species, and likely all bioluminescent beetles, use the same substrates beetle luciferin, ATP and oxygen to produce light. It is the structure of the particular luciferase enzyme that is the key to determining the color of the emitted light. We report here the molecular cloning of the P. scintillans luc gene and the expression and characterization of the corresponding novel recombinant luciferase enzyme. A comparison of the amino acid sequence with that of the highly similar P. pyralis enzyme and subsequent mutagenesis studies revealed that the single conservative amino acid change tyrosine to phenylalanine at position 255 accounted for the entire emission color difference. Additional mutagenesis and crystallographic studies were performed on a H-bond network, which includes the position 255 residue and five other stringently conserved beetle luciferase residues, that is proximal to the substrate/emitter binding site. The results are interpreted in the context of a speculative proposal that this network is key to the understanding of bioluminescence color determination.
- Subjects :
- 0301 basic medicine
Luminescence
Protein Conformation
Color
Molecular cloning
010402 general chemistry
Crystallography, X-Ray
01 natural sciences
Biochemistry
03 medical and health sciences
Luciferases, Firefly
Photinus pyralis
Bioluminescence
Animals
Luciferase
Physical and Theoretical Chemistry
Cloning, Molecular
Peptide sequence
Pyralis
biology
Fireflies
Hydrogen Bonding
General Medicine
biology.organism_classification
Luciferin
Recombinant Proteins
0104 chemical sciences
030104 developmental biology
Amino Acid Substitution
Mutagenesis, Site-Directed
Photinus
Subjects
Details
- ISSN :
- 17511097
- Volume :
- 93
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Photochemistry and photobiology
- Accession number :
- edsair.doi.dedup.....6aaf3d3589cfa157bff414b3f174d282