Continuous Assay for Acid Phosphatase Using Phenyl Phosphate

Redox activation of mitochondrial intermembrane space Cu,Zn-superoxide dismutase

“…The purity of isolated mitochondria was characterized enzymatically by measuring, in both granule-and Percoll-purified mitochondrial fractions, the activities of acid phosphatase (a lysosomal marker), urate oxidase (a peroxisomal marker), and fumarase and sulphite oxidase (mitochondrial markers), as described previously [19][20][21][22]. The activities of the marker enzymes in the mitochondrial fraction were expressed as percentages of total activities observed in the granule fraction.…”

Vibrational spectroscopic studies and ab initio calculations of phenyl phosphate disodium salt

“…[9,10] Phenyl phosphate has been used as the acid phosphatase substrate for about 50 years. [11] Nonlinear optics deals with the interaction of applied electromagnetic fields in various materials to generate new electromagnetic fields, altered in wavenumber, phase or other physical properties. [12] Organic molecules able to manipulate photonic signals efficiently are of importance in technologies such as optical communication, optical computing and dynamic image processing.…”

Positive cooperativity in substrate binding of human prostatic acid phosphatase entrapped in AOT–isooctane–water reverse micelles

“…The experiments have been carried out over the w 0 range 5 to 35, at constant overall enzyme concentration (9 nM), in pH 5.6 and 3.8, above and below the isoelectric point of PAP (equal 4.9 [3]). The previously prepared spectrophotometric methods for continuous monitoring of the reaction product have been used for initial reaction rate estimations [43,44]. In this paper reverse micelles have been chosen as microreactors of the precisely designed water-pool dimensions and have been employed to promote dissociation of dimeric PAP molecule to monomers at low w 0 or to promote association of dimeric PAP molecule to tetramers (or higher oligomers) at high w 0 .…”

“…The assay was conducted immediately at 20 • C by continuous monitoring of the absorbance of 1-naphthol at 322 nm ( ε = 2800 M −1 cm −1 ) [44] or of phenol at 278 nm (ε = 1500 M −1 cm −1 ) [43]. The initial reaction rate was calculated using Gillford spectrophotometer software.…”

“…For this purpose previously prepared spectrophotometric continuous acid phosphatase assays with 1-naphthyl phosphate and phenyl phosphate as substrates, usable in aqueous and micellar solutions, have been employed [43,44]. In order to determine steady-state reaction constants the experimental data were fitted to Hill rate equation.…”