A radiochemical assay for beta-ureidopropionase using radiolabeled N-carbamyl-beta-alanine obtained via hydrolysis of [2-(14)C]5, 6-dihydrouracil
| Autor: | A. B. P. Van Kuilenburg, A. H. van Gennip, H. van Lenthe |
|---|---|
| Přispěvatelé: | Other departments |
| Jazyk: | angličtina |
| Rok vydání: | 1999 |
| Předmět: |
Radioisotope Dilution Technique
Beta-ureidopropionase Coefficient of variation Kinetics Biophysics Sensitivity and Specificity Biochemistry Amidohydrolases chemistry.chemical_compound Hydrolysis Humans Carbon Radioisotopes Uracil Molecular Biology chemistry.chemical_classification Detection limit Chromatography Liquid scintillation counting Reproducibility of Results Dihydrouracil Cell Biology Carbon Dioxide Enzyme Liver chemistry beta-Alanine Scintillation Counting |
| Zdroj: | Analytical biochemistry, 272(2), 250-253. Academic Press Inc. |
| ISSN: | 0003-2697 |
| DOI: | 10.1006/abio.1999.4181 |
| Popis: | A radiochemical assay was developed to measure the activity of beta-ureidopropionase in human liver homogenates which is based on the detection of the reaction product (14)CO(2) by liquid scintillation counting. Radiolabeled N-carbamyl-beta-alanine was prepared within 15 min by a simple hydrolysis of [2-(14)C]5, 6-dihydrouracil under alkaline conditions at 37 degrees C. The enzymatic reaction proved to be linear with time up to at least 3.5 h and protein concentrations up to at least 1 mg/ml. Human beta-ureidopropionase obeyed Michaelis-Menten kinetics with an apparent Km for N-carbamyl-beta-alanine of 15.5 +/- 1.9 microM. The assay proved to be very accurate and sensitive with an intraassay coefficient of variation of 2% and a detection limit of 28 pmol for the product CO(2). |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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