Journal
ANALYTICAL CHEMISTRY
Volume 87, Issue 1, Pages 816-820Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac503958r
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Funding
- National Basic Research Program of China [2010CB732403]
- NSFC for Excellent Youth Scholars of China [21222506]
- NSFC [21175024, 21375021, 21375019]
- program for New Century Excellent Talents in University [NCET-12-0619]
- Nature Sciences Funding of Fujian Province [2014J06005]
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A fluorometric method for pyrophosphatase (PPase) activity detection was developed based on click chemistry. Cu(II) can coordinate with pyrophosphate (PPi), the addition of pyrophosphatase (PPase) into the above system can destroy the coordinate compound because PPase catalyzes the hydrolysis of PPi into inorganic phosphate and produces free Cu(II), and free Cu(II) can be reduced by sodium ascorbate (SA) to form Cu(I), which in turn initiates the ligating reaction between nonfluorescent 3-azidocoumarins and terminal alkynes to produce a highly fluorescent triazole complex, based on which, a simple and sensitive turn on fluorometric method for PPase can be developed. The fluorescence intensity of the system has a linear relationship with the logarithm of the PPase concentration in the range of 0.5 and 10 mU with a detection limit down to 0.2 mU (S/N = 3). This method is cost-effective and convenient without any labels or complicated operations. The proposed system was applied to screen the potential PPase inhibitor with high efficiency. The proposed method can be applied to diagnosis of PPase-related diseases.
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