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核心蛋白质组介导的治疗靶标挖掘和幽门螺杆菌的多表位疫苗设计。

Core proteome mediated therapeutic target mining and multi-epitope vaccine design for Helicobacter pylori.

机构信息

H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.

Department of Biochemistry, Abdul Wali Khan University Mardan, Mardan 23200, KP, Pakistan.

出版信息

Genomics. 2020 Sep;112(5):3473-3483. doi: 10.1016/j.ygeno.2020.06.026. Epub 2020 Jun 17.

Abstract

Helicobacter pylori is a Gram-negative spiral-shaped bacterium that infects half of the human population worldwide and causes chronic inflammation. In the present study, we used the art of computational biology for therapeutic drug targets identification and a multi-epitope vaccine against multi-strains of H. pylori. For drug target identification, we used different tools and softwares to identify human non-homologous but pathogen essential proteins, with virulent properties and involved in unique metabolic pathways of H. pylori. For this purpose, the core proteome of 84 strains of H. pylori was retrieved from EDGAR 2.3 database. There were 59,808 proteins sequences in these strains. Duplicates and paralogous protein sequence removal was followed by human non-homologous protein miningPathogen essential and virulent proteins were subjected to pathway analysis Subcellular localization of the virulent proteins was predicted and druggability was also checked, leading to 30 druggable targets based on their similarity with the approved drug targets in Drugbank. For immunoinformatics analysis, we selected two outer membrane proteins (HPAKL86_RS06305 and HPSNT_RS00950) and subjected to determined immunogenic B and T-Cell epitopes. The B and T-Cell overlapped epitopes were selected to design 9 different vaccine constructs by using linkers and adjuvants. Least allergenic and most antigenic construct (C-8) was selected as a promiscuous vaccine to elicit host immune response. Cloning and in silico expression of the constructed vaccine (C-8) was done to produce a clone having the desired (gene) vaccine construct. In conclusion, the prioritized therapeutic targets for 84 strains of H.pylori will be useful for future therapy design. Vaccine design may also prove useful in the quest for targeting multi-strains of H. pylori in patients.

摘要

幽门螺杆菌是一种革兰氏阴性螺旋形细菌,它感染了全球一半的人口,并导致慢性炎症。在本研究中,我们使用计算生物学技术来识别治疗药物靶点,并针对多种幽门螺杆菌菌株设计一种多表位疫苗。为了识别药物靶点,我们使用不同的工具和软件来识别人类非同源但病原体必需的蛋白质,这些蛋白质具有毒力特性,并参与幽门螺杆菌独特的代谢途径。为此,从 EDGAR 2.3 数据库中检索了 84 株幽门螺杆菌的核心蛋白质组。这些菌株中有 59808 个蛋白质序列。去除重复和旁系同源蛋白序列后,对人类非同源蛋白进行挖掘,然后对病原体必需和毒力蛋白进行途径分析。预测毒力蛋白的亚细胞定位,并检查其成药性,从而根据其与 Drugbank 中已批准药物靶点的相似性,确定了 30 个可成药的靶点。对于免疫信息学分析,我们选择了两种外膜蛋白(HPAKL86_RS06305 和 HPSNT_RS00950),并对其进行了确定的免疫原性 B 和 T 细胞表位分析。选择 B 和 T 细胞重叠表位,通过接头和佐剂设计 9 种不同的疫苗构建体。选择最小变应原性和最大抗原性构建体(C-8)作为一种混杂疫苗,以激发宿主免疫反应。构建疫苗(C-8)的克隆和计算机表达用于产生具有所需(基因)疫苗构建体的克隆。总之,针对 84 株幽门螺杆菌的优先治疗靶点将有助于未来的治疗设计。疫苗设计也可能有助于针对患者中的多种幽门螺杆菌菌株进行靶向治疗。

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