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• W2029584311 startingPage "4884" @default.
• W2029584311 abstract "A spectrophotometric method for the measurement of inorganic phosphate (P(i)) has been developed by using 2-amino-6-mercapto-7-methylpurine ribonucleoside and purine-nucleoside phosphorylase (purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1). This substrate gives an absorbance increase at 360 nm on phosphorolysis at pH 6.5-8.5, and at pH 7.6 the change in extinction coefficient is 11,000 M-1.cm-1. The Michaelis-Menten constants of the two substrates with the enzyme are 70 microM for the nucleoside and 26 microM for P(i); the kcat is 40 s-1 (25 degrees C). The assay was shown to quantitate P(i) in solution at concentrations at least down to 2 microM. It can be used to measure the kinetics of P(i) release from phosphatases, such as GTPases and ATPases, by coupling the two enzymic reactions. The utility of this assay was shown by three test systems: glycerol kinase plus D-glyceraldehyde acting as an ATPase and actin-activated myosin ATPase, and myosin subfragment 1, hydrolyzing a single turnover of ATP, releasing P(i) with a rate constant the same as the steady-state ATPase activity." @default.
• W2029584311 created "2016-06-24" @default.
• W2029584311 creator A5078281981 @default.
• W2029584311 date "1992-06-01" @default.
• W2029584311 modified "2023-10-02" @default.
• W2029584311 title "A continuous spectrophotometric assay for inorganic phosphate and for measuring phosphate release kinetics in biological systems." @default.
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• W2029584311 doi "https://doi.org/10.1073/pnas.89.11.4884" @default.
• W2029584311 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/49192" @default.
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