Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform

Ryo Nishihara, Emi Hoshino, Yoshiki Kakudate, Satoshi Kishigami, Naoko Iwasawa, Shin Ichi Sasaki, Takahiro Nakajima, Moritoshi Sato, Shigeru Nishiyama, Daniel Citterio, Koji Suzuki, Sung Bae Kim

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Native coelenterazine (nCTZ) is a common substrate to most marine luciferases and photoproteins. In this study, nine novel dye- and azide-conjugated CTZ analogues were synthesized by conjugating a series of fluorescent dyes or an azide group to the C-2 or C-6 position of the nCTZ backbone to obtain bulkiness-driven substrate specificity and potential chemiluminescence/bioluminescence resonance energy transfer (C/BRET). The investigation on the optical properties revealed that azide-conjugated CTZs emit greatly biased bioluminescence to ALucs and ca. 130 nm blue-shifted bioluminescence with RLuc8.6 in living animal cells or lysates. The corresponding kinetic study explains that azide-conjugated CTZ exerts higher catalytic efficiency than nCTZ. Nile red-conjugated CTZ completely showed red-shifted CRET spectra and characteristic BRET spectra with artificial luciferase 16 (ALuc16). No or less spectral overlap occurs among [Furimazine-NanoLuc], [6-N3-CTZ-ALuc26], [6-pi-OH-CTZ-RLuc8.6], and [6-N3-CTZ-RLuc8.6] pairs, because of the substrate-driven luciferase specificity and color shifts, providing a crosstalk-free multiplex bioassay platform. The unique bioluminescence system appends a new toolbox to bioassays and multiplex molecular imaging platforms. This study is the first example that systematically synthesized fluorescent dye-conjugated CTZs and applied them for a bioluminescence assay system.

Original languageEnglish
Pages (from-to)1922-1931
Number of pages10
JournalBioconjugate Chemistry
Volume29
Issue number6
DOIs
Publication statusPublished - 2018 Jun 20

Fingerprint

Bioluminescence
Molecular imaging
Molecular Imaging
Azides
Coloring Agents
Dyes
Luciferases
Fluorescent Dyes
Biological Assay
Bioassay
Luminescent Proteins
Substrates
Energy Transfer
Substrate Specificity
Luminescence
Chemiluminescence
Crosstalk
Color
Energy transfer
Assays

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Nishihara, R., Hoshino, E., Kakudate, Y., Kishigami, S., Iwasawa, N., Sasaki, S. I., ... Kim, S. B. (2018). Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform. Bioconjugate Chemistry, 29(6), 1922-1931. https://doi.org/10.1021/acs.bioconjchem.8b00188

Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform. / Nishihara, Ryo; Hoshino, Emi; Kakudate, Yoshiki; Kishigami, Satoshi; Iwasawa, Naoko; Sasaki, Shin Ichi; Nakajima, Takahiro; Sato, Moritoshi; Nishiyama, Shigeru; Citterio, Daniel; Suzuki, Koji; Kim, Sung Bae.

In: Bioconjugate Chemistry, Vol. 29, No. 6, 20.06.2018, p. 1922-1931.

Research output: Contribution to journalArticle

Nishihara, R, Hoshino, E, Kakudate, Y, Kishigami, S, Iwasawa, N, Sasaki, SI, Nakajima, T, Sato, M, Nishiyama, S, Citterio, D, Suzuki, K & Kim, SB 2018, 'Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform', Bioconjugate Chemistry, vol. 29, no. 6, pp. 1922-1931. https://doi.org/10.1021/acs.bioconjchem.8b00188
Nishihara R, Hoshino E, Kakudate Y, Kishigami S, Iwasawa N, Sasaki SI et al. Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform. Bioconjugate Chemistry. 2018 Jun 20;29(6):1922-1931. https://doi.org/10.1021/acs.bioconjchem.8b00188
Nishihara, Ryo ; Hoshino, Emi ; Kakudate, Yoshiki ; Kishigami, Satoshi ; Iwasawa, Naoko ; Sasaki, Shin Ichi ; Nakajima, Takahiro ; Sato, Moritoshi ; Nishiyama, Shigeru ; Citterio, Daniel ; Suzuki, Koji ; Kim, Sung Bae. / Azide- and Dye-Conjugated Coelenterazine Analogues for a Multiplex Molecular Imaging Platform. In: Bioconjugate Chemistry. 2018 ; Vol. 29, No. 6. pp. 1922-1931.
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AU - Sasaki, Shin Ichi

AU - Nakajima, Takahiro

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