Journal
JOURNAL OF BIOMOLECULAR SCREENING
Volume 18, Issue 8, Pages 879-889Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057113489731
Keywords
kynurenine; 3-hydroxykynurenine; kynurenine 3-monooxygenase; LC/MS/MS; microglia; human PBMCs
Funding
- CHDI Foundation
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Kynurenine monooxygenase (KMO) catalyzes the conversion of kynurenine to 3-hydroxykynurenine. Modulation of KMO activity has been implicated in several neurodegenerative diseases, including Huntington disease. Our goal is to develop potent and selective small-molecule KMO inhibitors with suitable pharmacokinetic characteristics for in vivo proof-of-concept studies and subsequent clinical development. We developed a comprehensive panel of biochemical and cell-based assays that use liquid chromatography/tandem mass spectrometry to quantify unlabeled kynurenine and 3-hydroxykynurenine. We describe assays to measure KMO inhibition in cell and tissue extracts, as well as cellular assays including heterologous cell lines and primary rat microglia and human peripheral blood mononuclear cells.
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