Hariprasad Gururao, Kota Divya, Baskar Singh Sundararajan, Srinivasan Alagiri, Adhikary Souparno
Department of Biophysics, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029 India.
Indian J Clin Biochem. 2014 Oct;29(4):430-41. doi: 10.1007/s12291-013-0377-1. Epub 2013 Sep 11.
Clonorchis sinensis or the Chinese liver fluke is one of the most prevalent parasites affecting a major population in the oriental countries. The parasite lacks lipid generating mechanisms but is exposed to fatty acid rich bile in the liver. A secretory phospholipase A2, an enzyme that breaks down complex lipids, is important for the growth of the parasite. The enzyme is also implicated in the pathogenesis leading up to the hepatic fibrosis and its complications including cancer. The five isoforms of this particular enzyme from the parasite therefore qualify as potential drug targets. In this study, a detailed structural and ligand binding analysis of the isoforms has been done by modeling. The overall three dimensional structures of the isoforms are well conserved with three helices and a β-wing stabilized by four disulfide bonds. There are characteristic differences at the calcium binding loop, hydrophobic channel and the C-terminal domain that can potentially be exploited for drug binding. But the most significant feature pertains to the catalytic site where the isoforms exhibit three variations of either a histidine-aspartate-tyrosine or histidine-glutamate-tyrosine or histidine-aspartate-phenylalanine. Molecular docking studies show that isoform specific residues and their conformations in the substrate binding hydrophobic channel make unique interactions with certain inhibitor molecules resulting in a perfect tight fit. The proposed ligand molecules have a predicted affinity in micro-molar to nano-molar range. Interestingly, few of the ligand binding interaction patterns is in accordance to the phylogenetic studies to thereby establish the usefulness of evolutionary mechanisms in aiding ligand design. The molecular diversity of the parasitic PLA2 described in this study provides a platform for personalized medicine in the therapeutics of clonorchiasis.
华支睾吸虫,即中华肝吸虫,是影响东方国家众多人口的最常见寄生虫之一。这种寄生虫缺乏脂质生成机制,但却暴露于肝脏中富含脂肪酸的胆汁中。一种分泌型磷脂酶A2,一种分解复杂脂质的酶,对该寄生虫的生长很重要。该酶还与导致肝纤维化及其并发症(包括癌症)的发病机制有关。因此,来自该寄生虫的这种特定酶的五种同工型有资格成为潜在的药物靶点。在本研究中,通过建模对这些同工型进行了详细的结构和配体结合分析。这些同工型的整体三维结构高度保守,有三个螺旋和一个由四个二硫键稳定的β翼。在钙结合环、疏水通道和C端结构域存在特征性差异,这些差异可能被用于药物结合。但最显著的特征与催化位点有关,在该位点,同工型表现出组氨酸-天冬氨酸-酪氨酸或组氨酸-谷氨酸-酪氨酸或组氨酸-天冬氨酸-苯丙氨酸的三种变体。分子对接研究表明,底物结合疏水通道中同工型特异性残基及其构象与某些抑制剂分子产生独特的相互作用,从而实现完美紧密结合。所提出的配体分子预测亲和力在微摩尔到纳摩尔范围内。有趣的是,一些配体结合相互作用模式与系统发育研究一致,从而确立了进化机制在辅助配体设计中的有用性。本研究中描述的寄生性磷脂酶A2的分子多样性为华支睾吸虫病治疗中的个性化医疗提供了一个平台。