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Impact of site-directed mutant luciferase on quantitative green and orange/red emission intensities in firefly bioluminescence.

Wang Y, Akiyama H, Terakado K, Nakatsu T - Sci Rep (2013)

Bottom Line: While the mutation caused different emission spectra, the spectra differed only in the intensity of the green component (λmax ~ 560 nm).In contrast, the orange (λmax ~ 610 nm) and red (λmax ~ 650 nm) components present in all the spectra were almost unaffected by the modifications to the luciferases and changes in pH.Our results reveal that the intensity of the green component is the unique factor that is influenced by the luciferase structure and other reaction conditions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Solid State Physics, University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 2778584, Japan. wang@issp.u-tokyo.ac.jp

ABSTRACT
Firefly bioluminescence has attracted great interest because of its high quantum yield and intriguing modifiable colours. Modifications to the structure of the enzyme luciferase can change the emission colour of firefly bioluminescence, and the mechanism of the colour change has been intensively studied by biochemists, structural biologists, optical physicists, and quantum-chemistry theorists. Here, we report on the quantitative spectra of firefly bioluminescence catalysed by wild-type and four site-directed mutant luciferases. While the mutation caused different emission spectra, the spectra differed only in the intensity of the green component (λmax ~ 560 nm). In contrast, the orange (λmax ~ 610 nm) and red (λmax ~ 650 nm) components present in all the spectra were almost unaffected by the modifications to the luciferases and changes in pH. Our results reveal that the intensity of the green component is the unique factor that is influenced by the luciferase structure and other reaction conditions.

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Related in: MedlinePlus

Photographs of bioluminescence catalyzed by wild-type (WT), Y257F, Y257A, Y257E, and Y257R mutant of Lcr luciferase at pH 8.0.The corresponding structures were generated from the crystal structure of Lcr luciferase according to ref. 12 (code 2D1R in Protein Data Bank) with PyMOL14.
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f1: Photographs of bioluminescence catalyzed by wild-type (WT), Y257F, Y257A, Y257E, and Y257R mutant of Lcr luciferase at pH 8.0.The corresponding structures were generated from the crystal structure of Lcr luciferase according to ref. 12 (code 2D1R in Protein Data Bank) with PyMOL14.

Mentions: Figure 1 shows photographs of the bioluminescence catalyzed by the wild-type (WT), Y257F, Y257A, Y257E, and Y257R Lcr luciferases at pH 8.0, the colours of which are yellow-green, yellow, orange, red, and yellow, respectively. Figure 2 (a) shows the quantitative spectra of bioluminescence catalyzed by wild-type (WT) Lcr luciferase and the four mutants Y257F/A/E/R at various pH values. The absolute spectra show all the photons emitted by consuming one mole of LH2. The quantum yields and spectral peak wavelengths (λmax) at pH 8.0 are summarized in Table 1.


Impact of site-directed mutant luciferase on quantitative green and orange/red emission intensities in firefly bioluminescence.

Wang Y, Akiyama H, Terakado K, Nakatsu T - Sci Rep (2013)

Photographs of bioluminescence catalyzed by wild-type (WT), Y257F, Y257A, Y257E, and Y257R mutant of Lcr luciferase at pH 8.0.The corresponding structures were generated from the crystal structure of Lcr luciferase according to ref. 12 (code 2D1R in Protein Data Bank) with PyMOL14.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750616&req=5

f1: Photographs of bioluminescence catalyzed by wild-type (WT), Y257F, Y257A, Y257E, and Y257R mutant of Lcr luciferase at pH 8.0.The corresponding structures were generated from the crystal structure of Lcr luciferase according to ref. 12 (code 2D1R in Protein Data Bank) with PyMOL14.
Mentions: Figure 1 shows photographs of the bioluminescence catalyzed by the wild-type (WT), Y257F, Y257A, Y257E, and Y257R Lcr luciferases at pH 8.0, the colours of which are yellow-green, yellow, orange, red, and yellow, respectively. Figure 2 (a) shows the quantitative spectra of bioluminescence catalyzed by wild-type (WT) Lcr luciferase and the four mutants Y257F/A/E/R at various pH values. The absolute spectra show all the photons emitted by consuming one mole of LH2. The quantum yields and spectral peak wavelengths (λmax) at pH 8.0 are summarized in Table 1.

Bottom Line: While the mutation caused different emission spectra, the spectra differed only in the intensity of the green component (λmax ~ 560 nm).In contrast, the orange (λmax ~ 610 nm) and red (λmax ~ 650 nm) components present in all the spectra were almost unaffected by the modifications to the luciferases and changes in pH.Our results reveal that the intensity of the green component is the unique factor that is influenced by the luciferase structure and other reaction conditions.

View Article: PubMed Central - PubMed

Affiliation: Institute for Solid State Physics, University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 2778584, Japan. wang@issp.u-tokyo.ac.jp

ABSTRACT
Firefly bioluminescence has attracted great interest because of its high quantum yield and intriguing modifiable colours. Modifications to the structure of the enzyme luciferase can change the emission colour of firefly bioluminescence, and the mechanism of the colour change has been intensively studied by biochemists, structural biologists, optical physicists, and quantum-chemistry theorists. Here, we report on the quantitative spectra of firefly bioluminescence catalysed by wild-type and four site-directed mutant luciferases. While the mutation caused different emission spectra, the spectra differed only in the intensity of the green component (λmax ~ 560 nm). In contrast, the orange (λmax ~ 610 nm) and red (λmax ~ 650 nm) components present in all the spectra were almost unaffected by the modifications to the luciferases and changes in pH. Our results reveal that the intensity of the green component is the unique factor that is influenced by the luciferase structure and other reaction conditions.

Show MeSH
Related in: MedlinePlus