Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition-Reference-Cited by-同舟云学术

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

Published:2023-12-22 Issue: Volume: Page:
ISSN:2198-3844
Container-title:Advanced Science
language:en
Short-container-title:Advanced Science

Author:

Ukegawa Tatsuya¹,Komatsu Toru¹^ORCID,Minoda Mayano¹,Matsumoto Takuya¹,Iwasaka Takumi¹,Mizuno Tadahaya¹,Tachibana Ryo¹,Sakamoto Shingo¹,Hanaoka Kenjiro²,Kusuhara Hiroyuki¹,Honda Kazufumi³⁴,Watanabe Rikiya⁵,Urano Yasuteru¹⁶

Affiliation:

1. Graduate School of Pharmaceutical Sciences The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 Japan

2. Graduate School of Pharmaceutical Sciences Keio University 1‐5‐30, Shibakoen, Minato‐ku Tokyo 105–8512 Japan

3. Graduate School of Medicine Nippon Medical School 1‐1‐5 Sendagi, Bunkyo‐ku Tokyo 113–8602 Japan

4. Institute for Advanced Medical Science Nippon Medical School 1‐1‐5 Sendagi, Bunkyo‐ku Tokyo 113–8602 Japan

5. Cluster for Pioneering Research Riken, 2‐1 Hirosawa, Wako Saitama 351‐0198 Japan

6. Graduate School of Medicine The University of Tokyo 7‐3‐1 Hongo, Bunkyo‐ku Tokyo 113‐0033 Japan

Abstract

AbstractSingle‐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.

Funder

Ministry of Education, Culture, Sports, Science and Technology

Naito Foundation

Mochida Memorial Foundation for Medical and Pharmaceutical Research

Publisher

Wiley

Subject

General Physics and Astronomy,General Engineering,Biochemistry, Genetics and Molecular Biology (miscellaneous),General Materials Science,General Chemical Engineering,Medicine (miscellaneous)

Link

https://onlinelibrary.wiley.com/doi/pdf/10.1002/advs.202306559

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3. Research Highlights

4. Microfabricated arrays of femtoliter chambers allow single molecule enzymology