Sarma Abhijit, Gunasekaran Dhandapani, Rex Devasahayam Arokia Balaya, Sikha Thoduvayil, Phukan Homen, Kiran Kumar Mangalaparthi, Pinto Sneha M, Prasad Thottethodi Subrahmanya Keshava, Madanan Madathiparambil G
Regional Medical Research Centre Port Blair, Indian Council of Medical Research, Port Blair 744103, Andaman and Nicobar Islands, India.
Department of Chemical Sciences, Ariel University, Ariel 70400, Israel.
Pathogens. 2021 Jul 6;10(7):852. doi: 10.3390/pathogens10070852.
Leptospirosis is a re-emerging form of zoonosis that is caused by the spirochete pathogen . Extracellular proteins play critical roles in the pathogenicity and survival of this pathogen in the host and environment. Extraction and analysis of extracellular proteins is a difficult task due to the abundance of enrichments like serum and bovine serum albumin in the culture medium, as is distinguishing them from the cellular proteins that may reach the analyte during extraction. In this study, extracellular proteins were separated as secretory proteins from the culture supernatant and surface proteins were separated during the washing of the cell pellet. The proteins identified were sorted based on the proportion of the cellular fractions and the extracellular fractions. The results showed the identification of 56 extracellular proteins, out of which 19 were exclusively extracellular. For those proteins, the difference in quantity with respect to their presence within the cell was found to be up to 1770-fold. Further, bioinformatics analysis elucidated characteristics and functions of the identified proteins. Orthologs of extracellular proteins in various species were found to be closely related among different pathogenic forms. In addition to the identification of extracellular proteins, this study put forward a method for the extraction and identification of extracellular proteins.
钩端螺旋体病是一种由螺旋体病原体引起的再度出现的人畜共患病形式。细胞外蛋白在该病原体在宿主和环境中的致病性及生存中起着关键作用。由于培养基中存在诸如血清和牛血清白蛋白等大量富集物,以及在提取过程中难以将细胞外蛋白与可能混入分析物中的细胞蛋白区分开来,细胞外蛋白的提取和分析是一项艰巨的任务。在本研究中,细胞外蛋白作为分泌蛋白从培养上清液中分离出来,而表面蛋白则在洗涤细胞沉淀时分离出来。鉴定出的蛋白质根据细胞组分和细胞外组分的比例进行分类。结果显示鉴定出56种细胞外蛋白,其中19种是仅存在于细胞外的。对于这些蛋白质,发现其在细胞内和细胞外的数量差异高达1770倍。此外,生物信息学分析阐明了鉴定出的蛋白质的特征和功能。发现不同致病形式的各种物种中细胞外蛋白的直系同源物密切相关。除了鉴定细胞外蛋白外,本研究还提出了一种细胞外蛋白的提取和鉴定方法。