Department of Molecular Parasitology, Sungkyunkwan University School of Medicine and Center for Molecular Medicine, Samsung Biomedical Research Center, 300 Cheoncheon-dong, Jangan-gu, Suwon, Gyeonggi-do 440-746, Republic of Korea.
Int J Parasitol. 2011 Sep;41(11):1207-15. doi: 10.1016/j.ijpara.2011.07.004. Epub 2011 Aug 7.
We previously identified a hydrophobic-ligand-binding protein (HLBP) of the Taenia solium metacestode (TsM), which might be involved in the uptake of fatty acids (FAs) from host environments. The TsM 150kDa HLBP was a hetero-oligomeric complex composed of multiple 7kDa (RS1) and 10kDa (CyDA, b1 and m13h) subunits, and displayed a wide spectrum of binding affinities toward various FA analogs. In this study, we analysed biochemical properties and phylogenetic relationships of the individual subunits. Despite the low sequence identity (average 26.5%), these subunit proteins conserved an α-helix-rich structural domain and the first introns inserted in each of the respective chromosomal genes were found to be orthologous to one another, suggesting their common evolutionary origin. The recombinant RS1 protein bound strongly to all of the FA analogs examined including 11-[(5-dimethylaminonaphthalene-1-sulfonyl)amino]undecanoic acid (DAUDA), but not to 16-(9-anthroyloxy)palmitic acid (16-AP). The interactive binding between RS1 and FA analogs was specifically interfered with by the addition of non-fluorescent FA molecules or antibodies specific to the 150kDa protein. Conversely, the 10kDa members reacted only with the palmitic acid-derived 16-AP, whose interactive force was strengthened by the presence of other FA molecules. The use of mutagenic RS1 proteins demonstrated that a structural/electrostatic integrity around the second α-helix, rather than the conventional Trp residue, was the major factor governing the hydrophobic interaction. The 7 and 10kDa proteins exhibited distinctive immunoreactive patterns against sera from neurocysticercosis patients. These collective data suggest that the paralogous protein family have gained diverse functions during their evolution, to ensure the maintenance of metabolic homeostasis and survival of TsMs in hostile host environments.
我们之前鉴定了猪带绦虫囊尾蚴(TsM)的一种疏水性配体结合蛋白(HLBP),它可能参与从宿主环境中摄取脂肪酸(FAs)。TsM 150kDa HLBP 是一种由多个 7kDa(RS1)和 10kDa(CyDA、b1 和 m13h)亚基组成的异源寡聚复合物,对各种 FA 类似物表现出广泛的结合亲和力。在这项研究中,我们分析了各个亚基的生化特性和系统发生关系。尽管这些亚基蛋白的序列同一性(平均为 26.5%)较低,但它们都保守着富含α-螺旋的结构域,并且在各自的染色体基因中插入的第一个内含子彼此同源,这表明它们具有共同的进化起源。重组 RS1 蛋白强烈结合我们研究的所有 FA 类似物,包括 11-[(5-二甲基氨基萘-1-磺酰基)氨基]十一烷酸(DAUDA),但不结合 16-(9-蒽酰氧基)棕榈酸(16-AP)。RS1 和 FA 类似物之间的相互作用结合受到非荧光 FA 分子或针对 150kDa 蛋白的特异性抗体的特异性干扰。相反,10kDa 成员仅与棕榈酸衍生的 16-AP 反应,其他 FA 分子的存在会增强其相互作用力。使用突变的 RS1 蛋白表明,第二α-螺旋周围的结构/静电完整性,而不是传统的色氨酸残基,是控制疏水性相互作用的主要因素。7 和 10kDa 蛋白对神经囊尾蚴病患者的血清表现出不同的免疫反应模式。这些综合数据表明,在进化过程中,这些平行蛋白家族获得了多样化的功能,以确保 TsM 在恶劣的宿主环境中维持代谢平衡和生存。