Dixit R, Das M, Seth P K, Mukhtar H
Environ Res. 1986 Aug;40(2):365-71. doi: 10.1016/s0013-9351(86)80111-6.
The binding of acrylamide (ACR) with purified bovine serum albumin (BSA) was studied. Binding of ACR with BSA was characterized by equilibrium dialysis, fluorescence studies, and ultraviolet spectroscopy. ACR was quantitated by high-pressure liquid chromatography. Equilibrium dialysis studies on the binding of ACR with BSA showed that more than 25% of the added ligand was bound to the protein at equilibrium. ACR produced a concentration-dependent decrease in uv absorbance of BSA, indicating reactivity of ACR with BSA. Fluorescence quenching studies of ACR binding showed a concentration-dependent quenching of the fluorescence of BSA. ACR also caused a concentration-dependent decrease in the fluorescence of sulfhydryl (-SH) groups present on BSA, implicating a role of protein -SH groups in the binding of ACR. Prior blocking of -SH groups by N-ethylmaleimide resulted in 16% inhibition of the initial binding of ACR, suggesting involvement of -SH groups. Our results demonstrate that ACR binds to BSA through both aromatic amino acids and -SH groups and that such binding may play an important role in pharmacodynamics and toxicity of ACR.
研究了丙烯酰胺（ACR）与纯化牛血清白蛋白（BSA）的结合情况。通过平衡透析、荧光研究和紫外光谱对ACR与BSA的结合进行了表征。采用高压液相色谱法对ACR进行定量分析。对ACR与BSA结合的平衡透析研究表明，在平衡状态下，超过25%的添加配体与蛋白质结合。ACR使BSA的紫外吸光度呈浓度依赖性降低，表明ACR与BSA发生了反应。对ACR结合的荧光猝灭研究表明，ACR对BSA的荧光有浓度依赖性猝灭作用。ACR还使BSA上存在的巯基（-SH）荧光呈浓度依赖性降低，这表明蛋白质的-SH基团在ACR结合中起作用。用N-乙基马来酰亚胺预先阻断-SH基团导致ACR初始结合受到16%的抑制，表明-SH基团参与其中。我们的结果表明，ACR通过芳香族氨基酸和-SH基团与BSA结合，并且这种结合可能在ACR的药效学和毒性中起重要作用。
