Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition
Abstract Single‐molecule enzyme activity assay is a platform that enables the analysis of enzyme activities at single proteoform level. The limitation of the targetable enzymes is the major drawback of the assay, but the general assay platform is reported to study single‐molecule enzyme activities o...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2024-03-01
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| Series: | Advanced Science |
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| Online Access: | https://doi.org/10.1002/advs.202306559 |
| _version_ | 1797263480832655360 |
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| author | Tatsuya Ukegawa Toru Komatsu Mayano Minoda Takuya Matsumoto Takumi Iwasaka Tadahaya Mizuno Ryo Tachibana Shingo Sakamoto Kenjiro Hanaoka Hiroyuki Kusuhara Kazufumi Honda Rikiya Watanabe Yasuteru Urano |
| author_facet | Tatsuya Ukegawa Toru Komatsu Mayano Minoda Takuya Matsumoto Takumi Iwasaka Tadahaya Mizuno Ryo Tachibana Shingo Sakamoto Kenjiro Hanaoka Hiroyuki Kusuhara Kazufumi Honda Rikiya Watanabe Yasuteru Urano |
| author_sort | Tatsuya Ukegawa |
| collection | DOAJ |
| description | Abstract Single‐molecule enzyme activity assay is a platform that enables the analysis of enzyme activities at single proteoform level. The limitation of the targetable enzymes is the major drawback of the assay, but the general assay platform is reported to study single‐molecule enzyme activities of esterases based on the coupled assay using thioesters as substrate analogues. The coupled assay is realized by developing highly water‐soluble thiol‐reacting probes based on phosphonate‐substituted boron dipyrromethene (BODIPY). The system enables the detection of cholinesterase activities in blood samples at single‐molecule level, and it is shown that the dissecting alterations of single‐molecule esterase activities can serve as an informative platform for activity‐based diagnosis. |
| first_indexed | 2024-04-25T00:13:41Z |
| format | Article |
| id | doaj.art-a9a6946f5e86432fbac5da93a75bd205 |
| institution | Directory Open Access Journal |
| issn | 2198-3844 |
| language | English |
| last_indexed | 2024-04-25T00:13:41Z |
| publishDate | 2024-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj.art-a9a6946f5e86432fbac5da93a75bd2052024-03-13T07:30:35ZengWileyAdvanced Science2198-38442024-03-011110n/an/a10.1002/advs.202306559Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate RecognitionTatsuya Ukegawa0Toru Komatsu1Mayano Minoda2Takuya Matsumoto3Takumi Iwasaka4Tadahaya Mizuno5Ryo Tachibana6Shingo Sakamoto7Kenjiro Hanaoka8Hiroyuki Kusuhara9Kazufumi Honda10Rikiya Watanabe11Yasuteru Urano12Graduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Pharmaceutical Sciences Keio University 1‐5‐30, Shibakoen, Minato‐ku Tokyo 105–8512 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanGraduate School of Medicine Nippon Medical School 1‐1‐5 Sendagi, Bunkyo‐ku Tokyo 113–8602 JapanCluster for Pioneering Research Riken, 2‐1 Hirosawa, Wako Saitama 351‐0198 JapanGraduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 JapanAbstract Single‐molecule enzyme activity assay is a platform that enables the analysis of enzyme activities at single proteoform level. The limitation of the targetable enzymes is the major drawback of the assay, but the general assay platform is reported to study single‐molecule enzyme activities of esterases based on the coupled assay using thioesters as substrate analogues. The coupled assay is realized by developing highly water‐soluble thiol‐reacting probes based on phosphonate‐substituted boron dipyrromethene (BODIPY). The system enables the detection of cholinesterase activities in blood samples at single‐molecule level, and it is shown that the dissecting alterations of single‐molecule esterase activities can serve as an informative platform for activity‐based diagnosis.https://doi.org/10.1002/advs.202306559chemical biologyenzymesenzymomicssingle‐molecule analysis |
| spellingShingle | Tatsuya Ukegawa Toru Komatsu Mayano Minoda Takuya Matsumoto Takumi Iwasaka Tadahaya Mizuno Ryo Tachibana Shingo Sakamoto Kenjiro Hanaoka Hiroyuki Kusuhara Kazufumi Honda Rikiya Watanabe Yasuteru Urano Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition Advanced Science chemical biology enzymes enzymomics single‐molecule analysis |
| title | Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition |
| title_full | Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition |
| title_fullStr | Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition |
| title_full_unstemmed | Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition |
| title_short | Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition |
| title_sort | thioester based coupled fluorogenic assays in microdevice for the detection of single molecule enzyme activities of esterases with specified substrate recognition |
| topic | chemical biology enzymes enzymomics single‐molecule analysis |
| url | https://doi.org/10.1002/advs.202306559 |
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