Amoebiasis, being endemic worldwide, is the second leading cause of parasite-associated morbidity and mortality after malaria. The human parasite Entamoeba histolytica is responsible for the disease. Metronidazole is considered as the gold standard for the treatment of amoebiasis, but this antibiotic is carcinogenic and the development of antibiotic resistance against E. histolytica is a major health concern. Chromosome segregation is irregular in this parasite due to the absence of a few cell cycle checkpoint proteins. Anaphase-promoting complex (APC/C or cyclosome) is an E3 ubiquitin ligase that synchronizes chromosome segregation and anaphase progression via the ubiquitin-proteasome system. Proteasome is considered to be an attractive drug target for protozoan parasites. For the present study, EhApc11 from E. histolytica, a homologue of Apc11 in humans, is selected for elucidating its structural and functional aspects by detailed in silico analysis and molecular methods. Its physicochemical characteristics, identification of probable interactors, 3D model and quality analysis are done using standard bioinformatics tools. cDNA sequence of EhAPC11 has been further cloned for molecular characterization. Conserved domain analysis revealed that EhApc11 belongs to the RING (really interesting new gene) superfamily and has ligand binding capacity. Expression study in Escherichia coli BL21 (DE3) revealed that the molecular weight of glutathione S-transferase (GST)-tagged protein is ~ 36 kDa. EhApc11 is a hydrophilic, thermostable, extracellular protein with potent antigenicity. The study will serve as a groundwork for future in-depth analysis regarding the validation of protein-protein interaction of EhApc11 with its substrates identified by STRING analysis and the potential of EhApc11 to serve as an anti-amoebic drug target.

中文翻译：

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来自人类病原体溶组织内阿米巴的后期促进复合物同系物的计算机分析和分子鉴定

阿米巴病在世界范围内流行，是仅次于疟疾的寄生虫相关发病率和死亡率的第二大原因。人类寄生虫溶组织内阿米巴是该疾病的罪魁祸首。甲硝唑被认为是治疗阿米巴病的金标准，但这种抗生素具有致癌性，并且对溶组织内阿米巴的抗生素耐药性的发展是一个主要的健康问题。由于缺乏一些细胞周期检查点蛋白，这种寄生虫的染色体分离是不规则的。后期促进复合物（APC/C 或环体）是一种 E3 泛素连接酶，通过泛素-蛋白酶体系统同步染色体分离和后期进展。蛋白酶体被认为是对原生动物寄生虫有吸引力的药物靶点。对于本研究，来自 E. histolytica 的 EhApc11，选择人类 Apc11 的同源物，通过详细的计算机分析和分子方法阐明其结构和功能方面。其理化特性、可能的相互作用物的识别、3D 模型和质量分析都是使用标准生物信息学工具完成的。EhAPC11 的 cDNA 序列已被进一步克隆用于分子表征。保守域分析表明，EhApc11 属于 RING（真正有趣的新基因）超家族，具有配体结合能力。在大肠杆菌 BL21 (DE3) 中的表达研究表明，谷胱甘肽 S-转移酶 (GST) 标记蛋白的分子量约为 36 kDa。EhApc11 是一种亲水、热稳定的细胞外蛋白，具有强抗原性。