Department of Chemistry, China Agricultural University, Beijing 100193, China.
Ecotoxicol Environ Saf. 2012 Apr;78:41-9. doi: 10.1016/j.ecoenv.2011.11.006. Epub 2012 Jan 10.
Malachite green is a triphenylmethane dye that is used extensively in many industrial and aquacultural processes, generating environmental concerns and health problems to human being. In this contribution, the complexation between lysozyme and malachite green was verified by means of computer-aided molecular modeling, steady state and time-resolved fluorescence, and circular dichroism (CD) approaches. The precise binding patch of malachite green in lysozyme has been identified from molecular modeling and ANS displacement, Trp-62, Trp-63, and Trp-108 residues of lysozyme were earmarked to possess high-affinity for this dye, the principal forces in the lysozyme-malachite green adduct are hydrophobic and π-π interactions. Steady state fluorescence proclaimed the complex of malachite green with lysozyme yields quenching through static type, which substantiates time-resolved fluorescence measurements that lysozyme-malachite green conjugation formation has an affinity of 10(3)M(-1). Moreover, via molecular modeling and also CD data, we can safely arrive at a conclusion that the polypeptide chain of lysozyme partially destabilized upon complexation with malachite green. The data emerged here will help to further understand the toxicological action of malachite green in human body.
孔雀石绿是一种三苯甲烷染料，广泛应用于许多工业和水产养殖过程中，引起了人们对环境和健康问题的关注。在本研究中，通过计算机辅助分子建模、稳态和时间分辨荧光以及圆二色性（CD）方法验证了溶菌酶与孔雀石绿的络合作用。从分子建模和 ANS 置换中确定了溶菌酶中孔雀石绿的精确结合位，溶菌酶的色氨酸 62、63 和 108 残基被指定为对这种染料具有高亲和力，溶菌酶-孔雀石绿加合物的主要力是疏水和π-π相互作用。稳态荧光表明，孔雀石绿与溶菌酶的复合物通过静态类型产生猝灭，这证实了时间分辨荧光测量结果，即溶菌酶-孔雀石绿缀合形成具有 10(3)M(-1)的亲和力。此外，通过分子建模和 CD 数据，我们可以得出结论，在与孔雀石绿络合后，溶菌酶的多肽链部分失稳。这里出现的数据将有助于进一步了解孔雀石绿在人体内的毒理学作用。
