University of Iowa, Iowa City, 52242-1112, United States.
Chem Biol Interact. 2011 Feb 1;189(3):153-60. doi: 10.1016/j.cbi.2010.11.009. Epub 2010 Dec 3.
Hydroxylated metabolites of polychlorinated biphenyls (OHPCBs) interact with rat sulfotransferase 1A1 (rSULT1A1) as substrates and inhibitors. Previous studies have shown that there are complex and incompletely understood structure-activity relationships governing the interaction of rSULT1A1 with these molecules. Furthermore, modification of the enzyme with glutathione disulfide (GSSG) results in the conversion of some OHPCBs from inhibitors to substrates. We have now examined estimated values for the acid-dissociation constant (K(a)) and the octanol-water distribution coefficient (D), as well as experimentally determined dissociation constants for enzyme complexes, to assist in the prediction of interactions of OHPCBs with rSULT1A1. Under reducing conditions, initial velocities for rSULT1A1-catalyzed sulfation exhibited a positive correlation with pK(a) and a negative correlation with logD of the OHPCBs. IC(50) values of inhibitory OHPCBs decreased with decreasing pK(a) values for both the glutathione (GSH)-pretreated and GSSG-pretreated forms of rSULT1A1. Comparison of GSH- and GSSG-pretreated forms of rSULT1A1 with respect to binding of OHPCB in the presence and absence of adenosine 3',5'-diphosphate (PAP) revealed that the dissociation constants with the two redox states of the enzyme were similar for each OHPCB. Thus, pK(a) and logD values are useful in predicting the binding of OHPCBs to the two redox forms of rSULT1A1 as well as the rates of sulfation of those OHPCBs that are substrates. However, the differences in substrate specificity for OHPCBs that are seen with changes in redox status of the enzyme are not directly related to specific structural effects of individual OHPCBs within inhibitory enzyme-PAP-OHPCB complexes.
多氯联苯的羟基代谢物(OHPCBs)作为底物和抑制剂与大鼠磺基转移酶 1A1(rSULT1A1)相互作用。先前的研究表明,rSULT1A1 与这些分子相互作用的结构-活性关系复杂且不完全清楚。此外,用谷胱甘肽二硫化物(GSSG)修饰酶会导致一些 OHPCBs 从抑制剂转变为底物。我们现在检查了酸离解常数(K(a))和辛醇-水分配系数(D)的估计值,以及酶复合物的实验测定解离常数,以帮助预测 OHPCBs 与 rSULT1A1 的相互作用。在还原条件下,rSULT1A1 催化的磺化初始速度与 OHPCB 的 pK(a)呈正相关,与 logD 呈负相关。对于 GSH-预处理和 GSSG-预处理形式的 rSULT1A1,抑制性 OHPCB 的 IC(50)值随 pK(a)值的降低而降低。与 GSH 和 GSSG 预处理形式的 rSULT1A1 相比,在存在和不存在腺苷 3',5'-二磷酸(PAP)的情况下,OHPCB 的结合表明,两种氧化还原状态下酶的解离常数相似。因此,pK(a)和 logD 值可用于预测 OHPCB 与 rSULT1A1 的两种氧化还原形式的结合,以及作为底物的那些 OHPCB 的磺化速率。然而,在酶的氧化还原状态变化时观察到的 OHPCB 的底物特异性差异与抑制酶-PAP-OHPCB 复合物中单个 OHPCB 的特定结构效应没有直接关系。