Department of Marine Science, Bharathidasan University, Tiruchirapalli, 620024, India.
Centre for Computational Biology and Bioinformatics, School of Computational and Integrative Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.
Interdiscip Sci. 2015 Sep;7(3):326-33. doi: 10.1007/s12539-015-0270-0. Epub 2015 Aug 8.
The Legionella pneumophila (Lp), human pathogen, causes severe and often fatal Legionnaires' disease, produces a major virulence factor, termed 'macrophage infectivity potentiator protein' (Mip), that is necessary for optimal multiplication of the bacteria within human alveolar macrophages. Mip exhibits peptidyl prolyl cis-trans isomerase (PPIase) activity, which can be inhibited by rapamycin and FK506. It was previously shown that substitutions at the catalytic residues, aspartate-142 position replaced to leucine-142 and tyrosine-185 position replaced to alanine-185 strongly reduces the PPIase activity of Mip proteins. Therefore, we aim to develop an in silico mutagenesis model for both important catalytic residues, validated the stability of the mutated model. Further, we have docked the known inhibitor rapamycin with Lp Mip (native) and mutants (D142L and Y185A) to analyze the conformational and binding mode. Electrostatic contributions and van der Waals interactions are the major driving forces for rapamycin binding and largely responsible for the binding differences between the Lp Mip (native and mutated) proteins.
嗜肺军团菌(Lp)是一种人类病原体,可引起严重且常致命的军团病,产生一种主要的毒力因子,称为“巨噬细胞感染增强蛋白”(Mip),该因子对于在人肺泡巨噬细胞内细菌的最佳繁殖是必需的。Mip 具有肽基脯氨酰顺反式异构酶(PPIase)活性,可被雷帕霉素和 FK506 抑制。先前的研究表明,催化残基上的取代,天冬氨酸-142 位置被亮氨酸-142 取代,酪氨酸-185 位置被丙氨酸-185 取代,强烈降低了 Mip 蛋白的 PPIase 活性。因此,我们旨在为这两个重要的催化残基开发一个计算机诱变模型,并验证突变模型的稳定性。此外,我们还用已知抑制剂雷帕霉素对接 Lp Mip(天然)和突变体(D142L 和 Y185A),以分析构象和结合模式。静电贡献和范德华相互作用是雷帕霉素结合的主要驱动力,并在很大程度上导致 Lp Mip(天然和突变体)蛋白之间的结合差异。