Continuous assay for acid phosphatase using 1-naphthyl phosphate as a substrate
| Autor: | Ewa Luchter-Wasylewska |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Male
Inorganic chemistry Phosphatase Acid Phosphatase Naphthols Naphthalenes Micelle General Biochemistry Genetics and Molecular Biology Substrate Specificity chemistry.chemical_compound Hydrolysis Organophosphorus Compounds Humans Enzyme kinetics Micelles Aqueous solution Chromatography biology Chemistry Acid phosphatase Prostate Substrate (chemistry) Hydrogen-Ion Concentration Phosphate Solutions Kinetics biology.protein Spectrophotometry Ultraviolet |
| Zdroj: | Acta biochimica Polonica. 44(4) |
| ISSN: | 0001-527X |
| Popis: | The described continuous acid phosphatase assay is based on kinetics of the release of 1-naphthol in the course of the enzyme-catalyzed hydrolysis of 1-naphthyl phosphate, measured at 320 nm in aqueous solution and at 322 nm in sodium-bis(2-ethylhexyl)sulfosuccinate isooctane-water reverse micelles in a broad pH range (1.0-8.2). The method allows precise determination of the initial rate of the reaction and therefore may be used in the steady-state and pre-steady-state studies on the phosphatase-catalyzed reaction. The kinetic parameters (Km and kcat) for human prostatic acid phosphatase in aqueous solution and in reverse micelles, at pH 3.8, 4.5 and 5.7, by the proposed 1-naphthyl phosphate assay have been determined. |
| Databáze: | OpenAIRE |
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