Kasap Murat, Karadenizli Aynur, Akpınar Gürler, Uzuner Hüseyin, Ayimugu Abula, Karaosmanoğlu Kübra, Er Doğanhan Kadir
Medical Biology Department/DEKART Proteomics Laboratory, Kocaeli University Medical Faculty, Umuttepe, Kocaeli, 41380, Turkey.
Department of Medical Microbiology, Kocaeli University Medical Faculty, Umuttepe, Kocaeli, 41380, Turkey.
Curr Microbiol. 2017 Feb;74(2):230-238. doi: 10.1007/s00284-016-1178-6. Epub 2016 Dec 19.
Francisella tularensis is the causative agent of tularemia. Although major contributors and the main mechanism of the virulence are well known, some of the molecular details are still missing. Proteomics studies regarding F. tularensis have provided snapshot pictures of the organism grown under different culture conditions to understand the mechanism of virulence. In general, such studies were carried out with standard strains e.g., LVS and did not involve comparisons of F. tularensis isolates from either clinical or environmental sources. In this study, we performed two-dimensional gel electrophoresis (2DE)-based proteomic analysis and compared the protein profiles of the F. tularensis subsp. holarctica strains isolated from the clinical and the environmental samples. Regulations were detected in 14 spots when twofold regulation criteria were applied. The regulated protein spots were subjected to MALDI-TOF/TOF analysis and identified. Classification of the identified proteins based on metabolic functions revealed that the translation machinery was the most varying metabolic processes among the isolates. Using normalized protein spot intensities, PCA analysis was also performed. The results indicated that the strain isolated from water source was different then the strains isolated from the patients. Most interestingly, the isolates were strikingly distinguishable from the standard NCTC 10857 strain.
土拉弗朗西斯菌是兔热病的病原体。尽管其毒力的主要贡献因素和主要机制已为人熟知，但一些分子细节仍不清楚。关于土拉弗朗西斯菌的蛋白质组学研究提供了该生物体在不同培养条件下生长的快照，以了解其毒力机制。一般来说，此类研究是使用标准菌株（如LVS）进行的，并未涉及对来自临床或环境来源的土拉弗朗西斯菌分离株的比较。在本研究中，我们进行了基于二维凝胶电泳（2DE）的蛋白质组学分析，并比较了从临床和环境样本中分离的土拉弗朗西斯菌亚种全北区菌株的蛋白质谱。当应用两倍调控标准时，在14个斑点中检测到了调控。对调控的蛋白质斑点进行了基质辅助激光解吸电离飞行时间/串联飞行时间（MALDI-TOF/TOF）分析并进行了鉴定。根据代谢功能对鉴定出的蛋白质进行分类显示，翻译机制是分离株中变化最大的代谢过程。使用标准化的蛋白质斑点强度，还进行了主成分分析（PCA）。结果表明，从水源分离的菌株与从患者分离的菌株不同。最有趣的是，这些分离株与标准的NCTC 10857菌株明显不同。
