Biochemistry Department, Tehran University of Medical Sciences, Tehran, Iran.
Gene. 2012 Aug 15;505(1):19-22. doi: 10.1016/j.gene.2012.05.057. Epub 2012 Jun 6.
GPx1 is one of the most important enzymes involved in oxidative balance so that, we studied the phenotype and genotype relationship of GPx1 activity and rs 1800668 (C/T) site and also evaluated the changes of GPx1 kinetic parameters in the rs 1800668 homozygotes. One hundred fifty eight subjects were recruited after clinical exams. The rs 1800668 (C/T) genotype distribution was identified using RFLP-PCR method. The hemolysate GPx1 activity was spectrophotometrically measured in a reaction coupled with glutathione reductase (GR). The GPx1 enzyme was purified using gel filtration chromatography with Sephacryl S-300 column and, Km(app) was studied in the rs 1800668 TT and CC homozygotes. The results showed that the GPx1 activity is significantly associated to the rs 1800668 (C/T) genotype distribution (P<0.05) so that, the GPx1 activity was high among the CC homozygotes (P<0.03). In addition, Km(app) for TBHP substrate in the TT homozygote (8.48 μM) was higher than the CC homozygote (5.74 μM). We concluded that the C allele within rs 1800668 position is related to the GPx1 activity and may be a potential factor involved in development of inflammatory events.
GPx1 是参与氧化平衡的最重要的酶之一，因此，我们研究了 GPx1 活性和 rs1800668（C/T）位点的表型和基因型关系，并评估了 rs1800668 纯合子中 GPx1 动力学参数的变化。在临床检查后，招募了 158 名受试者。使用 RFLP-PCR 方法确定 rs1800668（C/T）基因型分布。用分光光度法在与谷胱甘肽还原酶（GR）偶联的反应中测量溶血 GPx1 活性。使用 Sephacryl S-300 柱凝胶过滤色谱法纯化 GPx1 酶，并在 rs1800668 TT 和 CC 纯合子中研究 Km(app)。结果表明，GPx1 活性与 rs1800668（C/T）基因型分布显著相关（P<0.05），因此，CC 纯合子中的 GPx1 活性较高（P<0.03）。此外，TBHP 底物在 TT 纯合子（8.48 μM）中的 Km(app)高于 CC 纯合子（5.74 μM）。我们得出结论，rs1800668 位置的 C 等位基因与 GPx1 活性相关，可能是参与炎症事件发展的潜在因素。
