Department of Medical Microbiology & Parasitology, School of Medical Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
Gut Pathog. 2013 Dec 11;5(1):38. doi: 10.1186/1757-4749-5-38.
Bacteria exist widely in a diversity of natural environments. In order to survive adverse conditions such as nutrient depletion, biochemical and biological disturbances, and high temperature, bacteria have developed a wide variety of coping mechanisms. Temperature is one of the most important factors that can enhance the expression of microbial proteins. This study was conducted to investigate how outer membrane proteins (OMPs) of the bacterium Shigella flexneri respond to stress, especially during fever when the host's body temperature is elevated.
OMPs of S. flexneri ATCC 12022 and clinical isolate SH057 were extracted from an overnight culture grown at 37, 38.5, and 40°C. Comparisons of the expressed proteins under the different growth conditions were based on equal numbers of bacterial cells loaded in the SDS-PAGE gels. Separated proteins were stained with Coomassie brilliant blue. Selected proteins showing increased expression at 38.5 and 40°C were characterized by performing MALDI-ToF-ToF.
Different degrees of expression were demonstrated for different proteins expressed at 37°C compared to 38.5 and 40°C. The proteins with molecular sizes of 18.4, 25.6, and 57.0 kDa showed increased expression level at increasing temperature and were identified as Dps, WrbA, and PepA, respectively.
This study revealed that strains of S. flexneri respond at the proteomic level during stress caused by elevated temperature by decreasing the expression of proteins, maintaining the level of important proteins, or enhancing the levels of proteins presumably involved in survival and virulence.
细菌广泛存在于各种自然环境中。为了应对营养物质枯竭、生化和生物干扰以及高温等不利条件,细菌发展出了各种各样的应对机制。温度是增强微生物蛋白表达的最重要因素之一。本研究旨在探讨福氏志贺菌的外膜蛋白(OMPs)如何应对压力,特别是在宿主体温升高引起发热时。
从在 37°C、38.5°C 和 40°C 下培养过夜的细菌中提取福氏志贺菌 ATCC 12022 和临床分离株 SH057 的 OMPs。基于加载在 SDS-PAGE 凝胶中的等量细菌细胞,比较不同生长条件下表达的蛋白质。用考马斯亮蓝染色分离的蛋白质。对在 38.5°C 和 40°C 下表达增加的选定蛋白质进行 MALDI-ToF-ToF 分析。
与 37°C 相比,不同蛋白质在 38.5°C 和 40°C 下的表达程度不同。分子量为 18.4、25.6 和 57.0 kDa 的蛋白质在温度升高时表达水平增加,分别被鉴定为 Dps、WrbA 和 PepA。
本研究揭示了福氏志贺菌株在高温引起的应激中,通过降低蛋白质的表达、维持重要蛋白质的水平或增强可能参与生存和毒力的蛋白质水平,在蛋白质组水平上作出反应。