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Single-Molecule Oxidoreductase Activity Analysis for Activity-Based Diagnosis Based on Proteoform Alterations

Mayano Minoda, Junpei Hatakeyama, N Nagano, Tadahaya Mizuno, T. Iwasaka, Seiichi Shiga, Kazuki Takahashi, Hideto Hiraide, Shingo Sakamoto, Yu Kagami, Ayumi Kashiro, Kazufumi Honda, Yuki Sugiura, Kazuhiko Mishima, Masayo Mishima, Hiroyuki Kusuhara, Yasuteru Urano, Toru Komatsu

2025Journal of the American Chemical Society11 citationsDOI

Abstract

-dependent oxidoreductases, leveraging a new NAD(P)H-responsive fluorogenic probe optimized for microdevice-based fluorometric detection. This platform enabled the detection of enzyme activities in blood and cerebrospinal fluid (CSF), including lactate dehydrogenase, glucose-6-phosphate dehydrogenase, and hexokinases. We demonstrate its potential for activity-based diagnosis by detecting altered populations of enzyme activity species in blood and CSF from liver damage in brain tumor patients.

Topics & Concepts

ChemistryNAD+ kinaseEnzymeLactate dehydrogenaseOxidoreductaseBiochemistryEnzyme assayCerebrospinal fluidDehydrogenasePathologyMedicineMitochondrial Function and PathologyGlioma Diagnosis and TreatmentCancer, Hypoxia, and Metabolism
Single-Molecule Oxidoreductase Activity Analysis for Activity-Based Diagnosis Based on Proteoform Alterations | Litcius