Demir Dudu, Gencer Nahit, Arslan Oktay
a Department of Agricultural Biotechnology, Faculty of Agriculture , Suleyman Demirel University , Isparta , Turkey and.
b Department of Chemistry, Faculty of Art and Science , Balikesir University , Balikesir , Turkey.
J Enzyme Inhib Med Chem. 2016;31(2):247-52. doi: 10.3109/14756366.2015.1018242. Epub 2015 Mar 20.
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.
在本研究中，开发了一种用于人对氧磷酶1（hPON1）的替代纯化方法，该方法采用两步程序，即硫酸铵沉淀和琼脂糖-4B-L-酪氨酸-3-氨基菲疏水相互作用色谱法。该酶的SDS-聚丙烯酰胺凝胶电泳显示出一条表观分子量为43 kDa的单一谱带。该酶被纯化了219倍，最终比活性为4,408,400 U/mg，产率为10%。此外，我们研究了一些合成代谢化合物，如玉米赤霉醇、17β-雌二醇、己烯雌酚、催产素和群勃龙对该酶活性的体外影响，以了解这些分子更好的抑制特性。这五种合成代谢化合物剂量依赖性地降低了hPON1的活性，抑制常数在毫摩尔-微摩尔范围内。结果表明，这些化合物在低浓度下对hPON1具有抑制作用，IC50值范围为0.064至16.900 µM。
