A rapid fluorometric assay for erythrocyte pyridoxal kinase
| Autor: | C.T. McQuilkin, C.U. Hicks, John A. Kark, R.D. Reynolds, M.J. Haut |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
Adult
Male Erythrocytes Biochemistry Hemolysis chemistry.chemical_compound medicine Humans Pyridoxal phosphate Tyrosine Pyridoxal kinase activity Pyridoxal Kinase Pyridoxal Phosphotransferases Pyridoxine Metabolism Pyridoxal kinase Kinetics Spectrometry Fluorescence chemistry Pyridoxal Phosphate Female Hemoglobin medicine.drug |
| Zdroj: | Biochemical medicine. 27(1) |
| ISSN: | 0006-2944 |
| Popis: | We describe a method for measurement of erythrocyte pyridoxal kinase activity in hemolysates, in which pyridoxal phosphate is oxidized and assayed by fluorometry. We also use this method to measure pyridoxal phosphate synthesis by intact erythrocytes suspended with pyridoxal or pyridoxine. The fluorometric method and the more laborious tyrosine apodecarboxylase method provide identical results. Binding of pyridoxal compounds to hemoglobin causes a small increase in Km for pyridoxal and loss of 5% of pyridoxal phosphate, but has no effect on the Vmax of pyridoxal kinase. The average coefficient of variance is 3% for hemolysate pyridoxal kinase activity and 10% for measurement of pyridoxal phosphate synthesis by intact erythrocytes. There is little increase in variance for observations spanning 18 to 24 months. The fluorometric method allows rapid, reproducible measurement of pyridoxal phosphate metabolism in hemolysates or intact erythrocytes. |
| Databáze: | OpenAIRE |
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