title: An alternative purification method for human serum paraoxonase 1 and its interaction with methidathion.
abstract: In this study, an alternative purification method for human Paraoxonase 1 (hPON1) enzyme was developed using two-step procedures, namely ammonium sulphate precipitation and Sepharose-4B-L-tyrosine-1-aminoanthracene hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis of the enzyme indicates a single band with an apparent MW of 43 kDa. The enzyme was purified 674-fold with a yield of 16%. Furthermore, we examined the in vitro effect of methidathion on the enzyme activity to understand the better inhibitory properties of the compound. Methidathion is a highly toxic insecticide used to control a broad spectrum of agricultural insect and mite pests. IC50value was found to be 0.130 mM for the pesticide. Methidathion showed a competitive inhibition with Ki of 0.119 mM for paraoxon.
authors.affiliation: a Department of Chemistry , Faculty of Art and Science, Balikesir University , Balikesir , Turkey.
authors.forename: Nahit
authors.initials: N
authors.lastname: Gençer
authors.affiliation: a Department of Chemistry , Faculty of Art and Science, Balikesir University , Balikesir , Turkey.
authors.forename: Emre
authors.initials: E
authors.lastname: Yavuz
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D043303:Aryldialkylphosphatase / Q000037:antagonists & inhibitors / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D001667:Binding, Competitive; D011232:Chemical Precipitation; D002845:Chromatography / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007306:Insecticides / Q000737:chemistry*; D007700:Kinetics; D008970:Molecular Weight; D009946:Organothiophosphorus Compounds / Q000737:chemistry*; D010261:Paraoxon / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D004791:Enzyme Inhibitors; D007306:Insecticides; D009946:Organothiophosphorus Compounds; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; D010261:Paraoxon; D000645:Ammonium Sulfate; C005828:methidathion
keywords: PON1; hydrophobic interaction chromatography; inhibition; methidathion; purification