Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green
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 c...
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MDPI AG
2020-03-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/5/1116 |
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| author | Nattha Yongwattana Nutsara Mekjinda Tulyapruek Tawonsawatruk Itaru Hamachi Akio Ojida Jirarut Wongkongkatep |
| author_facet | Nattha Yongwattana Nutsara Mekjinda Tulyapruek Tawonsawatruk Itaru Hamachi Akio Ojida Jirarut Wongkongkatep |
| author_sort | Nattha Yongwattana |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T05:16:34Z |
| format | Article |
| id | doaj.art-5b0e495f46f947b0988b3fef537c709a |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-04-13T05:16:34Z |
| publishDate | 2020-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-5b0e495f46f947b0988b3fef537c709a2022-12-22T03:00:53ZengMDPI AGMolecules1420-30492020-03-01255111610.3390/molecules25051116molecules25051116Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto GreenNattha Yongwattana0Nutsara Mekjinda1Tulyapruek Tawonsawatruk2Itaru Hamachi3Akio Ojida4Jirarut Wongkongkatep5Department of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, ThailandDepartment of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, ThailandDepartment of Orthopedics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, 270 Rama 6 Road, Bangkok 10400, ThailandDepartment of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, JapanGraduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, JapanDepartment of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, ThailandCalcium 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.https://www.mdpi.com/1420-3049/25/5/1116cppdpseudogoutenpp1ppaseapyrasesynovial fluidfluorescence detectionmolecular sensor |
| spellingShingle | Nattha Yongwattana Nutsara Mekjinda Tulyapruek Tawonsawatruk Itaru Hamachi Akio Ojida Jirarut Wongkongkatep Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green Molecules cppd pseudogout enpp1 ppase apyrase synovial fluid fluorescence detection molecular sensor |
| title | Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green |
| title_full | Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green |
| title_fullStr | Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green |
| title_full_unstemmed | Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green |
| title_short | Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green |
| title_sort | fluorescence differentiation of atp related multiple enzymatic activities in synovial fluid as a marker of calcium pyrophosphate deposition disease using kyoto green |
| topic | cppd pseudogout enpp1 ppase apyrase synovial fluid fluorescence detection molecular sensor |
| url | https://www.mdpi.com/1420-3049/25/5/1116 |
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