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Fluorescent and Colorimetric Dual-Readout Assay for Inorganic Pyrophosphatase with Cu2+-Triggered Oxidation of o-Phenylenediamine.
- Source :
-
Analytical Chemistry . 1/19/2016, Vol. 88 Issue 2, p1355-1361. 7p. - Publication Year :
- 2016
-
Abstract
- We demonstrate a rationally designed fluorescent and colorimetric dual-readout strategy for the highly sensitive, quantitative determination of inorganic pyrophosphatase (PPase) activity, a key hydrolytic enzyme involved in a variety of metabolic processes. Inspired by the selective oxidative and chromogenic reaction of o-phenylenediamine (OPD) with Cu2+, the special inhibitory effects of pyrophosphate (PPi) on the oxidative ability of Cu2+, and the specific hydrolysis of PPi into orthophosphate by PPase, a convenient small molecule OPD-based analytical system was developed for Cu2+/PPi recognition and PPase activity assay. We have confirmed that Cu2+ acts as the oxidant in the reaction and the main chromogenic product of OPD is 2,3-diaminophenazine (usually called OPDox) in the assay by combining the ESI-MS, 1H NMR, and XPS spectra analysis. Direct electrochemical insights into the Cu2+-triggered and PPi-inhibited mechanism were performed by cyclic voltammetry characterizations of the Cu2+ in the absence and presence of PPi for the first time. Furthermore, the proposed analytical system with clear response mechanism exhibits a promising outlook for the PPase activity assay in real biological samples and inhibitor screening. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00032700
- Volume :
- 88
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- Analytical Chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 112903533
- Full Text :
- https://doi.org/10.1021/acs.analchem.5b03848