Abstract
The present study demonstrates cation-driven optical properties of artificial luciferases (ALucs) from copepod luciferases, as an optical readout for bioanalysis. An assignment of the supersecondary structure code (SSC) of ALucs revealed that ALucs carry a helix-loop-helix structure, which appears at the same sites of the EF-hands of typical Ca2+-binding proteins. A mutagenesis study shows that the EF-hand-like structure is a pivotal site for enzymatic activity. The effects of 20 kinds of mono- and multivalent cations on ALuc activities were estimated with column-purified ALuc16. High pH values boost the ALuc activities with both the native coelenterazine and an analog called 6-pi-OH-CTZ. Multivalent cations, Ca(II), Mg(II), and Cr(VI), elevate and prolong the ALuc activities, whereas Co(II), Cu(II) and Pb(II) greatly hamper the ALuc activities. Ca(II) greatly prolongs the optical intensities, suggesting a contribution to the structural robustness of ALucs. The inhibitory effect of multivalent cations on the ALuc activities was utilized for creating dose-response curves. The intrinsic cation-driven selectivity and optical intensity of ALucs enable researchers to constitute de novo biosensors for multivalent cations.
| Original language | English |
|---|---|
| Pages (from-to) | 955-960 |
| Number of pages | 6 |
| Journal | Analytical Sciences |
| Volume | 31 |
| Issue number | 10 |
| Publication status | Published - 2015 |
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Keywords
- Artificial luciferase
- Bioluminescence
- Metal cation
- Supersecondary structure code
ASJC Scopus subject areas
- Analytical Chemistry
Cite this
Cation-driven optical properties of artificial luciferases. / Kim, Sung Bae; Miller, Simon; Suzuki, Nobuhiro; Senda, Toshiya; Nishihara, Ryo; Suzuki, Koji.
In: Analytical Sciences, Vol. 31, No. 10, 2015, p. 955-960.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Cation-driven optical properties of artificial luciferases
AU - Kim, Sung Bae
AU - Miller, Simon
AU - Suzuki, Nobuhiro
AU - Senda, Toshiya
AU - Nishihara, Ryo
AU - Suzuki, Koji
PY - 2015
Y1 - 2015
N2 - The present study demonstrates cation-driven optical properties of artificial luciferases (ALucs) from copepod luciferases, as an optical readout for bioanalysis. An assignment of the supersecondary structure code (SSC) of ALucs revealed that ALucs carry a helix-loop-helix structure, which appears at the same sites of the EF-hands of typical Ca2+-binding proteins. A mutagenesis study shows that the EF-hand-like structure is a pivotal site for enzymatic activity. The effects of 20 kinds of mono- and multivalent cations on ALuc activities were estimated with column-purified ALuc16. High pH values boost the ALuc activities with both the native coelenterazine and an analog called 6-pi-OH-CTZ. Multivalent cations, Ca(II), Mg(II), and Cr(VI), elevate and prolong the ALuc activities, whereas Co(II), Cu(II) and Pb(II) greatly hamper the ALuc activities. Ca(II) greatly prolongs the optical intensities, suggesting a contribution to the structural robustness of ALucs. The inhibitory effect of multivalent cations on the ALuc activities was utilized for creating dose-response curves. The intrinsic cation-driven selectivity and optical intensity of ALucs enable researchers to constitute de novo biosensors for multivalent cations.
AB - The present study demonstrates cation-driven optical properties of artificial luciferases (ALucs) from copepod luciferases, as an optical readout for bioanalysis. An assignment of the supersecondary structure code (SSC) of ALucs revealed that ALucs carry a helix-loop-helix structure, which appears at the same sites of the EF-hands of typical Ca2+-binding proteins. A mutagenesis study shows that the EF-hand-like structure is a pivotal site for enzymatic activity. The effects of 20 kinds of mono- and multivalent cations on ALuc activities were estimated with column-purified ALuc16. High pH values boost the ALuc activities with both the native coelenterazine and an analog called 6-pi-OH-CTZ. Multivalent cations, Ca(II), Mg(II), and Cr(VI), elevate and prolong the ALuc activities, whereas Co(II), Cu(II) and Pb(II) greatly hamper the ALuc activities. Ca(II) greatly prolongs the optical intensities, suggesting a contribution to the structural robustness of ALucs. The inhibitory effect of multivalent cations on the ALuc activities was utilized for creating dose-response curves. The intrinsic cation-driven selectivity and optical intensity of ALucs enable researchers to constitute de novo biosensors for multivalent cations.
KW - Artificial luciferase
KW - Bioluminescence
KW - Metal cation
KW - Supersecondary structure code
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UR - http://www.scopus.com/inward/citedby.url?scp=84947546266&partnerID=8YFLogxK
M3 - Article
C2 - 26460358
AN - SCOPUS:84947546266
VL - 31
SP - 955
EP - 960
JO - Analytical Sciences
JF - Analytical Sciences
SN - 0910-6340
IS - 10
ER -