Medical Microbiology and Molecular Biology Laboratory, Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, UP, 202002, India.
School of Computational & Integrative Sciences (SC&IS), Jawaharlal Nehru University, New Delhi, 110067, India.
Protein J. 2022 Feb;41(1):166-178. doi: 10.1007/s10930-021-10038-1. Epub 2022 Jan 6.
The aim of the present study, is to identify potential targets against the highly pathogenic bacteria Streptococcus mutans that causes dental caries as well as the deadly infection of endocarditis. The powerful and highly sensitive technique of liquid chromatography-mass spectrometry (LC-MS/MS) identified 321 proteins of S. mutans when grown under stressful conditions induced by the antibiotic bacitracin. These 321 proteins were subjected to the insilico method of subtractive proteomics to screen out potential targets by utilizing different analyses like CD-HIT, non-homologous sequence screening, KEGG pathway, essentiality screening, gut-flora non-homology, and codon usage analysis. A database of essential proteins was employed to find sequence homology of non-paralogous proteins to determine proteins which are essential for bacterial survival. Cellular localization analysis of the selected proteins was done to localize them inside the cell along with physico-chemical characterization and druggability analysis. Using computational tools, 22 proteins out of 321, that are functionally distinguishable from their human counterparts and passed the criterion of a potential therapeutic candidate were identified. The selected proteins comprise central energy metabolic proteins, virulence factors, proteins of the sortase family, and essentiality factors. The presented analyses identified proteins of the sortase family, which appear as key therapeutic targets against caries infection. These proteins regulate a number of virulence factors, thus can be simultaneously inhibited to obstruct multiple virulence pathways.
本研究旨在鉴定针对高致病性细菌变形链球菌(Streptococcus mutans)的潜在靶标,该细菌可引起龋齿以及致命的感染性心内膜炎。在抗生素杆菌肽诱导的应激条件下,强大而高度敏感的液相色谱-质谱联用(LC-MS/MS)技术鉴定了变形链球菌生长时的 321 种蛋白质。利用 CD-HIT、非同源序列筛选、KEGG 途径、必需性筛选、肠道菌群非同源性和密码子使用分析等不同分析方法,对这 321 种蛋白质进行了消减蛋白质组学的计算机方法筛选,以筛选出潜在的靶标。利用数据库中的必需蛋白质,寻找非旁系同源蛋白质的序列同源性,以确定对细菌生存至关重要的蛋白质。对选定蛋白质进行细胞定位分析,以确定它们在细胞内的位置,同时进行理化特性分析和可药性分析。使用计算工具,从 321 种蛋白质中鉴定出 22 种在功能上与人类蛋白质不同且符合潜在治疗候选物标准的蛋白质。选定的蛋白质包括中心能量代谢蛋白、毒力因子、天冬酰胺内肽酶家族的蛋白质和必需性因子。所提出的分析确定了天冬酰胺内肽酶家族的蛋白质,它们是抗龋齿感染的关键治疗靶标。这些蛋白质调节多种毒力因子,因此可以同时抑制以阻止多种毒力途径。