Development of red-shifted mutants derived from luciferase of Brazilian click beetle Pyrearinus termitilluminans

Tomoki Nishiguchi; Toshimichi Yamada; Yusuke Nasu; Mashiho Ito; Hideaki Yoshimura; Takeaki Ozawa

doi:10.1117/1.JBO.20.10.101205

27 August 2015 Development of red-shifted mutants derived from luciferase of Brazilian click beetle Pyrearinus termitilluminans

Tomoki Nishiguchi, Toshimichi Yamada, Yusuke Nasu, Mashiho Ito, Hideaki Yoshimura, Takeaki Ozawa

Author Affiliations +

Journal of Biomedical Optics, Vol. 20, Issue 10, 101205 (August 2015). https://doi.org/10.1117/1.JBO.20.10.101205

Abstract

Luciferase, a bioluminescent protein, has been used as an analytical tool to visualize intracellular phenomena. Luciferase with red light emission is particularly useful for bioluminescence imaging because of its high transmittance in mammalian tissues. However, the luminescence intensity of existing luciferases with their emission over 600 nm is insufficient for imaging studies because of their weak intensities. We developed mutants of Emerald luciferase (Eluc) from Brazilian click beetle (Pyrearinus termitilluminans), which emits the strongest bioluminescence among beetle luciferases. We successively introduced four amino acid mutations into the luciferase based on a predicted structure of Eluc using homology modeling. Results showed that quadruple mutations R214K/H241K/S246H/H347A into the beetle luciferase emit luminescence with emission maximum at 626 nm, 88-nm red-shift from the wild-type luciferase. This mutant luciferase is anticipated for application in in vivo multicolor imaging in living samples.

Citation Download Citation

Tomoki Nishiguchi, Toshimichi Yamada, Yusuke Nasu, Mashiho Ito, Hideaki Yoshimura, and Takeaki Ozawa "Development of red-shifted mutants derived from luciferase of Brazilian click beetle Pyrearinus termitilluminans," Journal of Biomedical Optics 20(10), 101205 (27 August 2015). https://doi.org/10.1117/1.JBO.20.10.101205

Published: 27 August 2015

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
7 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

CITATIONS

Cited by 20 scholarly publications and 4 patents.

Explore citations on Lens.org

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Bioluminescence

Luminescence

In vivo imaging

Proteins

Tissues

3D modeling

Visualization

Show All Keywords

RELATED CONTENT

Motion corrected photoacoustic difference imaging of fluorescent contrast agents
Proceedings of SPIE (March 15 2016)

Super-duper chemiluminescent proteins applicable to wide range of bioimaging
Proceedings of SPIE (February 08 2017)

Engineering of bacterial phytochromes for in vivo imaging (Conference Presentation)
Proceedings of SPIE (January 01 1900)

Identification and characterization of neutrophil extracellular trap shapes in flow...
Proceedings of SPIE (March 01 2017)

Visualization of tumor-induced VEGF expression using in vivo bioluminescence
Proceedings of SPIE (June 14 2004)

Photoacoustic imaging of the near infrared fluorescent protein iRFP in...
Proceedings of SPIE (February 23 2012)

Subscribe to Digital Library

Receive Erratum Email Alert

Show All Keywords

Keywords/Phrases

Search In:

Publication Years