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DMSO-tolerant ornithine decarboxylase (ODC) tandem assay optimised for high-throughput screening

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Title
DMSO-tolerant ornithine decarboxylase (ODC) tandem assay optimised for high-throughput screening
Author(s)
Mingu Gordon Park; Suyeon Yellena Kim; C. Justin Lee
Publication Date
2023-12
Journal
Journal of Enzyme Inhibition and Medicinal Chemistry, v.38, no.1, pp.309 - 318
Publisher
Taylor and Francis Ltd.
Abstract
© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.Ornithine decarboxylase (ODC), the first rate-limiting enzyme in polyamine synthesis, has emerged as a therapeutic target for cancer and Alzheimer’s disease (AD). To inhibit ODC, α-difluoromethylornithine (DFMO), an irreversible ODC inhibitor, has been widely used. However, due to its poor pharmacokinetics, the need for discovery of better ODC inhibitors is inevitable. For high-throughput screening (HTS) of ODC inhibitors, an ODC enzyme assay using supramolecular tandem assay has been introduced. Nevertheless, there has been no study utilising the ODC tandem assay for HTS, possibly due to its intolerability to dimethyl sulfoxide (DMSO), a common amphipathic solvent used for drug libraries. Here we report a DMSO-tolerant ODC tandem assay in which DMSO-dependent fluorescence quenching becomes negligible by separating enzyme reaction and putrescine detection. Furthermore, we optimised human cell-line-based mass production of ODC for HTS. Our newly developed assay can be a crucial first step in discovering more effective ODC modulators than DFMO.
URI
https://pr.ibs.re.kr/handle/8788114/12456
DOI
10.1080/14756366.2022.2150186
ISSN
1475-6366
Appears in Collections:
Center for Cognition and Sociality(인지 및 사회성 연구단) > 1. Journal Papers (저널논문)
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