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    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. © 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.

    Citation

    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 (Weinheim, Baden-Wurttemberg, Germany). 2024 Mar;11(10):e2306559

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    PMID: 38140707

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