title: An alternative purification method for human serum paraoxonase 1 and its interactions with anabolic compounds.
abstract: In this study, an alternative purification method for human paraoxonase 1 (hPON1) enzyme was developed using two-step procedures, namely, ammonium sulfate precipitation and Sepharose-4B-L-tyrosine-3-aminophenantrene hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis of the enzyme indicates a single band with an apparent M(W) of 43 kDa. The enzyme was purified 219-fold with a final specific activity of 4,408,400 U/mg and a yield of 10%. Furthermore, we examined the in vitro effects of some anabolic compounds, such as zeranol, 17 β-estradiol, diethylstilbestrol, oxytocin, and trenbolone on the enzyme activity to understand the better inhibitory properties of these molecules. The five anabolic compounds dose dependently decreased the activity of hPON1 with inhibition constants in the millimolar-micromolar range. The results show that these compounds exhibit inhibitory effects on hPON1 at low concentrations with IC50 values ranging from 0.064 to 16.900 µM.
authors: a Department of Agricultural Biotechnology, Faculty of Agriculture , Suleyman Demirel University , Isparta , Turkey and.
authors: Dudu
authors: D
authors: Demir
authors: b Department of Chemistry, Faculty of Art and Science , Balikesir University , Balikesir , Turkey.
authors: Nahit
authors: N
authors: Gencer
authors: b Department of Chemistry, Faculty of Art and Science , Balikesir University , Balikesir , Turkey.
authors: Oktay
authors: O
authors: Arslan
mesh_terms: D045930:Anabolic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D043303:Aryldialkylphosphatase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D002847:Chromatography, Agarose / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D020128:Inhibitory Concentration 50; D010121:Oxytocin / Q000494:pharmacology; D014204:Trenbolone Acetate / Q000494:pharmacology; D014443:Tyrosine / Q000737:chemistry; D015029:Zeranol / Q000494:pharmacology
chemical_list: D045930:Anabolic Agents; D014443:Tyrosine; D004958:Estradiol; D010121:Oxytocin; D015029:Zeranol; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human; D014204:Trenbolone Acetate
keywords: Anabolic compounds; hydrophobic interaction chromatography; inhibition; purification