Evaluation of a kinetic method for prostatic acid phosphatase with use of self-indicating substrate, 2,6-dichloro-4-nitrophenyl phosphate.
The purity, spectral characteristics, and rate of nonenzymatic hydrolysis of 2,6-dichloro-4-nitrophenyl phosphate (DCNPP) were determined. Rates of DCNPP hydrolysis by prostatic acid phosphatase (PAP) and erythrocytic acid phosphatase (EAP) (both EC 3.1.3.2) were measured in the absence and in the presence of various alcohols. 1.5-Pentanediol was the most effective transphosphorylation agent for specifically enhancing the activity of PAP. 1,4-Butanediol also enhanced PAP activity but markedly inhibited EAP activity. Bovine and human serum albumin preparations also accelerated the hydrolysis of DCNPP. DCNPP can be used for the continuous or multipoint-rate assay of PAP.
Valcour
A A
AA
Department of Pathology, Hartford Hospital, CT 06115.
Bowers
G N
GN
McComb
R B
RB
D000135:Acid Phosphatase / Q000378:metabolism*; D000438:Alcohols / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004789:Enzyme Activation; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D007700:Kinetics; D008297:Male; D009596:Nitrophenols / Q000378:metabolism; D000073718:Pancreatitis-Associated Proteins; D011467:Prostate / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet
D000438:Alcohols; D007202:Indicators and Reagents; D009596:Nitrophenols; D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; C054323:2,6-dichloro-4-nitrophenyl phosphate; D000135:Acid Phosphatase