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Kinetic Analysis of HIF Prolyl Hydroxylases
Peppi Koivunen1, Johanna Myllyharju2
1Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, Biocenter Oulu, University of Oulu, Oulu, Finland. peppi.karppinen@oulu.fi.
Abstract:
Kinetic analyses of HIF prolyl 4-hydroxylases (HIF-P4Hs) allow determination of substrate, cosubstrate and cofactor requirements, analysis of the reaction rate, and inhibitory properties of the isoenzymes in vitro. Here we describe an assay measuring the substrate hydroxylation-coupled decarboxylation of radioactive 2-oxoglutarate to radioactive carbon dioxide as a fast, efficient, and diverse method to analyze the enzyme kinetics of HIF-P4Hs.
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