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Article

Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green

1
Department of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, Thailand
2
Department of Orthopedics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, 270 Rama 6 Road, Bangkok 10400, Thailand
3
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan
4
Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
*
Author to whom correspondence should be addressed.
Molecules 2020, 25(5), 1116; https://doi.org/10.3390/molecules25051116
Received: 29 January 2020 / Revised: 24 February 2020 / Accepted: 25 February 2020 / Published: 2 March 2020
Calcium pyrophosphate deposition disease (CPPD) is a crystal induced inflammation in joints, and causes severe pain in elderly people. The accumulation of pyrophosphate (PPi) in synovial fluid (SF) results from several enzymatic reactions, especially the highly activated e-NPPs, which catalyze the conversion of ATP to PPi. This study demonstrates the detection of relative catalytic activity of 3 enzymes—ecto-nucleotide pyrophosphatase/phosphodiesterases (e-NPPs), tissue nonspecific alkaline phosphatase (TNAP), and ecto-nucleoside triphosphate diphosphohydrolases (e-NTPDases)—using a single molecular sensor called Kyoto Green. Kyoto Green exhibits excellent performance in sensing the catalytic activity of the commercial representatives of the e-NPPs, TNAP, and e-NTPDases, which are ENPP1, PPase, and apyrase, respectively, in both single-enzyme and multi-enzyme assays. Analysis of SF enzymes in 19 SF samples from human and swine revealed moderate activity of e-NPPs, high activity of e-NTPDases, and low activity of TNAP. Our newly developed method for analysis of multiple enzymatic activities using Kyoto Green in biological SF will assist improvement in accuracy of the CPPD prognosis/diagnosis, which will minimize unnecessary medical procedures. View Full-Text
Keywords: CPPD; pseudogout; ENPP1; PPase; apyrase; synovial fluid; fluorescence detection; molecular sensor CPPD; pseudogout; ENPP1; PPase; apyrase; synovial fluid; fluorescence detection; molecular sensor
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MDPI and ACS Style

Yongwattana, N.; Mekjinda, N.; Tawonsawatruk, T.; Hamachi, I.; Ojida, A.; Wongkongkatep, J. Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green. Molecules 2020, 25, 1116. https://doi.org/10.3390/molecules25051116

AMA Style

Yongwattana N, Mekjinda N, Tawonsawatruk T, Hamachi I, Ojida A, Wongkongkatep J. Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green. Molecules. 2020; 25(5):1116. https://doi.org/10.3390/molecules25051116

Chicago/Turabian Style

Yongwattana, Nattha, Nutsara Mekjinda, Tulyapruek Tawonsawatruk, Itaru Hamachi, Akio Ojida, and Jirarut Wongkongkatep. 2020. "Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green" Molecules 25, no. 5: 1116. https://doi.org/10.3390/molecules25051116

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