Fabrication of a new lineage of artificial luciferases from natural luciferase pools

Sung Bae Kim, Ryo Nishihara, Daniel Citterio, Koji Suzuki

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The fabrication of artificial luciferases (ALucs) with unique optical properties has a fundamental impact on bioassays and molecular imaging. In this study, we developed a new lineage of ALucs with unique substrate preferences by extracting consensus amino acids from the alignment of 25 copepod luciferase sequences available in natural luciferase pools. The primary sequence was first created with a sequence logo generator resulting in a total of 11 sibling sequences. Phylogenetic analysis shows that the newly fabricated ALucs form an independent branch, genetically isolated from the natural luciferases, and from a prior series of ALucs produced by our laboratory using a smaller basis set. The new lineage of ALucs were strongly luminescent in living mammalian cells with specific substrate selectivity to native coelenterazine. A single-residue-level comparison of the C-terminal sequences of new ALucs reveals that some amino acids in the C-terminal ends are greatly influential on the optical intensities but limited in the color variance. The success of this approach guides on how to engineer and functionalize marine luciferases for bioluminescence imaging and assays.

Original languageEnglish
Pages (from-to)594-599
Number of pages6
JournalACS Combinatorial Science
Volume19
Issue number9
DOIs
Publication statusPublished - 2017

Fingerprint

Luciferases
Fabrication
Bioluminescence
Molecular imaging
Amino Acids
Bioassay
Substrates
Assays
Optical properties
Cells
Color
Imaging techniques
Engineers

Keywords

  • Artificial luciferase
  • Bioluminescence
  • Coelenterazine
  • Copepod luciferase
  • Frequency analysis

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Fabrication of a new lineage of artificial luciferases from natural luciferase pools. / Kim, Sung Bae; Nishihara, Ryo; Citterio, Daniel; Suzuki, Koji.

In: ACS Combinatorial Science, Vol. 19, No. 9, 2017, p. 594-599.

Research output: Contribution to journalArticle

Kim, Sung Bae ; Nishihara, Ryo ; Citterio, Daniel ; Suzuki, Koji. / Fabrication of a new lineage of artificial luciferases from natural luciferase pools. In: ACS Combinatorial Science. 2017 ; Vol. 19, No. 9. pp. 594-599.
@article{970f43f4e64044e885a03b89ff95c7cb,
title = "Fabrication of a new lineage of artificial luciferases from natural luciferase pools",
abstract = "The fabrication of artificial luciferases (ALucs) with unique optical properties has a fundamental impact on bioassays and molecular imaging. In this study, we developed a new lineage of ALucs with unique substrate preferences by extracting consensus amino acids from the alignment of 25 copepod luciferase sequences available in natural luciferase pools. The primary sequence was first created with a sequence logo generator resulting in a total of 11 sibling sequences. Phylogenetic analysis shows that the newly fabricated ALucs form an independent branch, genetically isolated from the natural luciferases, and from a prior series of ALucs produced by our laboratory using a smaller basis set. The new lineage of ALucs were strongly luminescent in living mammalian cells with specific substrate selectivity to native coelenterazine. A single-residue-level comparison of the C-terminal sequences of new ALucs reveals that some amino acids in the C-terminal ends are greatly influential on the optical intensities but limited in the color variance. The success of this approach guides on how to engineer and functionalize marine luciferases for bioluminescence imaging and assays.",
keywords = "Artificial luciferase, Bioluminescence, Coelenterazine, Copepod luciferase, Frequency analysis",
author = "Kim, {Sung Bae} and Ryo Nishihara and Daniel Citterio and Koji Suzuki",
year = "2017",
doi = "10.1021/acscombsci.7b00081",
language = "English",
volume = "19",
pages = "594--599",
journal = "ACS Combinatorial Science",
issn = "2156-8952",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Fabrication of a new lineage of artificial luciferases from natural luciferase pools

AU - Kim, Sung Bae

AU - Nishihara, Ryo

AU - Citterio, Daniel

AU - Suzuki, Koji

PY - 2017

Y1 - 2017

N2 - The fabrication of artificial luciferases (ALucs) with unique optical properties has a fundamental impact on bioassays and molecular imaging. In this study, we developed a new lineage of ALucs with unique substrate preferences by extracting consensus amino acids from the alignment of 25 copepod luciferase sequences available in natural luciferase pools. The primary sequence was first created with a sequence logo generator resulting in a total of 11 sibling sequences. Phylogenetic analysis shows that the newly fabricated ALucs form an independent branch, genetically isolated from the natural luciferases, and from a prior series of ALucs produced by our laboratory using a smaller basis set. The new lineage of ALucs were strongly luminescent in living mammalian cells with specific substrate selectivity to native coelenterazine. A single-residue-level comparison of the C-terminal sequences of new ALucs reveals that some amino acids in the C-terminal ends are greatly influential on the optical intensities but limited in the color variance. The success of this approach guides on how to engineer and functionalize marine luciferases for bioluminescence imaging and assays.

AB - The fabrication of artificial luciferases (ALucs) with unique optical properties has a fundamental impact on bioassays and molecular imaging. In this study, we developed a new lineage of ALucs with unique substrate preferences by extracting consensus amino acids from the alignment of 25 copepod luciferase sequences available in natural luciferase pools. The primary sequence was first created with a sequence logo generator resulting in a total of 11 sibling sequences. Phylogenetic analysis shows that the newly fabricated ALucs form an independent branch, genetically isolated from the natural luciferases, and from a prior series of ALucs produced by our laboratory using a smaller basis set. The new lineage of ALucs were strongly luminescent in living mammalian cells with specific substrate selectivity to native coelenterazine. A single-residue-level comparison of the C-terminal sequences of new ALucs reveals that some amino acids in the C-terminal ends are greatly influential on the optical intensities but limited in the color variance. The success of this approach guides on how to engineer and functionalize marine luciferases for bioluminescence imaging and assays.

KW - Artificial luciferase

KW - Bioluminescence

KW - Coelenterazine

KW - Copepod luciferase

KW - Frequency analysis

UR - http://www.scopus.com/inward/record.url?scp=85032634513&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032634513&partnerID=8YFLogxK

U2 - 10.1021/acscombsci.7b00081

DO - 10.1021/acscombsci.7b00081

M3 - Article

C2 - 28742969

AN - SCOPUS:85032634513

VL - 19

SP - 594

EP - 599

JO - ACS Combinatorial Science

JF - ACS Combinatorial Science

SN - 2156-8952

IS - 9

ER -