Matsuhashi Takuto
| Institute for self-Evolving Smart Societies | Project/Specially Appointed Assistant Professor |
| Information Technology Center | Project/Specially Appointed Assistant Professor |
Researcher Information
Career
Research Activity Information
Paper
- Phrixotrix luciferase and 6′-aminoluciferins reveal a larger luciferin phenolate binding site and provide novel far-red combinations for bioimaging purposes
V. R. Bevilaqua; T. Matsuhashi; G. Oliveira; P. S. L. Oliveira; T. Hirano; V. R. Viviani
Scientific Reports, Springer Science and Business Media LLC, 9, 1, 21 Jun. 2019, Peer-reviwed, Abstract
How the unique luciferase of Phrixothrix hirtus (PxRE) railroad worm catalyzes the emission of red bioluminescence using the same luciferin of fireflies, remains a mystery. Although PxRE luciferase is a very attractive tool for bioanalysis and bioimaging in hemoglobin rich tissues, it displays lower quantum yield (15%) when compared to green emitting luciferases (>40%). To identify which parts of PxRE luciferin binding site (LBS) determine bioluminescence color, and to develop brighter and more red-shifted emitting luciferases, we compared the effects of site-directed mutagenesis and of larger 6′-substituted aminoluciferin analogues (6′-morpholino- and 6′-pyrrolidinyl-LH) on the bioluminescence properties of PxRE and green-yellow emitting beetle luciferases. The effects of mutations in the benzothiazolyl and thiazolyl parts of PxRE LBS on the KM and catalytic efficiencies, indicated their importance for luciferin binding and catalysis. However, the absence of effects on the bioluminescence spectrum indicated a less interactive LBS in PxRE during light emission. Mutations at the bottom of LBS of PxRE blue-shifted the spectra and increased catalytic efficiency, suggesting that lack of interactions of this part of LBS with excited oxyluciferin phenolate underlie red light emission. The much higher bioluminescence activity and red-shifted spectra of PxRE luciferase with 6′-morpholino- (634 nm) and 6′-pyrrolidinyl-luciferins (644 nm), when compared to other beetle luciferases, revealed a larger luciferin phenolate binding pocket. The size and orientation of the side-chains of L/I/H348 are critical for amino-analogues accommodation and modulate bioluminescence color, affecting the interactions and mobility of excited oxyluciferin phenolate. The PxRE luciferase and 6′-aminoluciferins provide potential far-red combinations for bioimaging applications.
Scientific journal - Electronic and Steric Effects of Cyclic Amino Substituents of Luciferin Analogues on a Firefly Luciferin–Luciferase Reaction
Michio Kakiuchi; Soichiro Ito; Masahiro Kiyama; Fumiya Goto; Takuto Matsuhashi; Minoru Yamaji; Shojiro Maki; Takashi Hirano
Chemistry Letters, Oxford University Press (OUP), 46, 8, 1090-1092, 05 Aug. 2017, Peer-reviwed
Scientific journal - Bioluminescence of Beetle Luciferases with 6′-Amino-
d -luciferin Analogues Reveals Excited Keto-oxyluciferin as the Emitter and Phenolate/Luciferin Binding Site Interactions Modulate Bioluminescence Colors
Vadim R. Viviani; Deimison Rodrigues Neves; Danilo Trabuco Amaral; Rogilene A. Prado; Takuto Matsuhashi; Takashi Hirano
Biochemistry, American Chemical Society (ACS), 53, 32, 5208-5220, 04 Aug. 2014, Peer-reviwed
Scientific journal - Spectroscopic studies of the color modulation mechanism of firefly (beetle) bioluminescence with amino-analogs of luciferin and oxyluciferin
Hirano, T; Nagai, H; Matsuhashi, T; Hasumi, Y; Iwano, S; Ito, K; Maki, S; Niwa, H; Viviani, V. R
Photochem. Photobiol. Sci., 11, 8, 1281-1284, Aug. 2012, Peer-reviwed
Scientific journal, English - Spectroscopic studies of the color modulation mechanism of firefly (beetle) bioluminescence with amino-analogs of luciferin and oxyluciferin
Takashi Hirano; Hiroyuki Nagai; Takuto Matsuhashi; Yosuke Hasumi; Satoshi Iwano; Kazuto Ito; Shojiro Maki; Haruki Niwa; Vadim R. Viviani
Photochemical and Photobiological Sciences, Springer Science and Business Media LLC, 11, 8, 1281-1284, 01 Aug. 2012, Peer-reviwed
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