Human liver paraoxonase (PON1): subcellular distribution and characterization.
The subcellular localization and different biochemical properties of a human hepatic microsomal enzyme that hydrolyses paraoxon (paraoxonase, PON1) were studied and compared to the paraoxon hydrolase activity found in human plasma as well as in rat liver and plasma. Having evaluated the influence of the postmortem interval by a parallel experiment performed in rats, we conclude that the paraoxonase activity was preferentially localized in the microsomal fraction. The enzyme reaction was optimized according to temperature, pH, buffer, ionic strength, substrate concentration, and enzyme protein concentration. The characterization of human liver paraoxonase included the study of optimum pH, pH stability, heat inactivation assays, and kinetic parameters (K(m) and Vmax). In addition, the enzyme activity showed an absolute requirement for exogenous calcium. The activity was lost after incubation with EDTA and partially restored by the addition of calcium; however, other metals assayed were not able to activate the human liver enzyme as did calcium. Our results support the possible identity between human plasma and liver paraoxonases. In spite of the technical difficulties of this study and the possible interference of the postmortem changes in the results, this article represents the first systematic approach to the characterization of human liver paraoxonase.
Gonzalvo
M C
MC
Department of Legal Medicine and Toxicology Service, Faculty of Medicine, University Hospital, University of Granada, Spain.
Gil
F
F
Hernandez
A F
AF
Rodrigo
L
L
Villanueva
E
E
Pla
A
A
D000818:Animals; D043303:Aryldialkylphosphatase; D002118:Calcium / Q000378:metabolism; D004950:Esterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008658:Inactivation, Metabolic; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions / Q000201:enzymology*
D004950:Esterases; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; D002118:Calcium