Thioester-Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single-Molecule Enzyme Activities of Esterases with Specified Substrate Recognition

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

研究成果: Article査読

抄録

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.

本文言語English
論文番号2306559
ジャーナルAdvanced Science
11
10
DOI
出版ステータスPublished - 2024 3月 13

ASJC Scopus subject areas

  • 医学(その他)
  • 化学工学一般
  • 材料科学一般
  • 生化学、遺伝学、分子生物学(その他)
  • 工学一般
  • 物理学および天文学一般

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