Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

Sun Sixiang; Kumi Y. Inoue; Shusaku Shiomoto; Shinichiro Takano; Kosuke Ino; Hitoshi Shiku; Tomokazu Matsue

doi:10.1002/celc.201600700

Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

Sun Sixiang, Kumi Y. Inoue, Shusaku Shiomoto, Shinichiro Takano, Kosuke Ino, Hitoshi Shiku, Tomokazu Matsue

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

Abstract

This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31–2.5×10⁻² units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.

Original language	English
Pages (from-to)	941-946
Number of pages	6
Journal	ChemElectroChem
Volume	4
Issue number	4
DOIs	https://doi.org/10.1002/celc.201600700
Publication status	Published - 2017 Apr 1

Keywords

amperometric detection
apoptosis
biosensing
caspase-3
peptide substrate

ASJC Scopus subject areas

Catalysis
Electrochemistry

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10.1002/celc.201600700

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title = "Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3",

abstract = "This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31–2.5×10−2 units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.",

keywords = "amperometric detection, apoptosis, biosensing, caspase-3, peptide substrate",

author = "Sun Sixiang and Inoue, {Kumi Y.} and Shusaku Shiomoto and Shinichiro Takano and Kosuke Ino and Hitoshi Shiku and Tomokazu Matsue",

note = "Funding Information: This research was supported by Grant-in-Aid for Young Scientists (B) (No. 23750073) from the Japan Society for the Promotion of Science (JSPS) and the Supporting Industry Project by the Ministry of Economy, Trade and Industry. This research was partly supported by a Grant-in-Aid for Scientific Research (A) (No. 16H02280) from JSPS, and by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program from the Japan Science and Technology Agency (JST). Publisher Copyright: {\textcopyright} 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim",

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T1 - Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

AU - Sixiang, Sun

AU - Inoue, Kumi Y.

AU - Shiomoto, Shusaku

AU - Takano, Shinichiro

AU - Ino, Kosuke

AU - Shiku, Hitoshi

AU - Matsue, Tomokazu

N1 - Funding Information: This research was supported by Grant-in-Aid for Young Scientists (B) (No. 23750073) from the Japan Society for the Promotion of Science (JSPS) and the Supporting Industry Project by the Ministry of Economy, Trade and Industry. This research was partly supported by a Grant-in-Aid for Scientific Research (A) (No. 16H02280) from JSPS, and by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program from the Japan Science and Technology Agency (JST). Publisher Copyright: © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

PY - 2017/4/1

Y1 - 2017/4/1

N2 - This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31–2.5×10−2 units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.

AB - This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31–2.5×10−2 units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.

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KW - peptide substrate

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Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

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