Universidade Federal do Rio de Janeiro, Departamento de Microbiologia Médica, Laboratório de Biologia de Anaeróbios, Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro, Instituto de Biofísica Carlos Chagas Filho e Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Bioimagens, Laboratório de Ultraestrutura Celular Hertha Meyer, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2022 Jun 17;117:e220035. doi: 10.1590/0074-02760220035. eCollection 2022.
Clostridioides difficile is the most common cause of nosocomial diarrhea associated with antibiotic use. The disease's symptoms are caused by enterotoxins, but other surface adhesion factors also play a role in the pathogenesis. These adhesins will bind to components of extracellular matrix.
There is a lack of knowledge on MSCRAMM, this work set-out to determine the adhesive properties of several C. difficile ribotypes (027, 133, 135, 014, 012) towards laminin-1 (LMN-1).
A binding experiment revealed that different ribotypes have distinct adhesion capabilities. To identify this adhesin, an affinity chromatography column containing LMN-1 was prepared and total protein extracts were analysed using mass spectrometry.
Strains from ribotypes 012 and 027 had the best adhesion when incubated with glucose supplementations (0.2%, 0.5%, and 1%), while RT135 had a poor adherence. The criteria were not met by RT014 and RT133. In the absence of glucose, there was no adhesion for any ribotype, implying that glucose is required and plays a significant role in adhesion.
These findings show that in the presence of glucose, each C. difficile ribotype interacts differently with LMN-1, and the adhesin responsible for recognition could be SlpA protein.
艰难梭菌是与抗生素使用相关的医院获得性腹泻的最常见原因。该疾病的症状是由肠毒素引起的,但其他表面粘附因子也在发病机制中起作用。这些粘附素将与细胞外基质的成分结合。
对 MSCRAMM 缺乏了解,本研究旨在确定几种艰难梭菌核糖体型(027、133、135、014、012)对层粘连蛋白-1(LMN-1)的粘附特性。
结合实验表明,不同的核糖体型具有不同的粘附能力。为了鉴定这种粘附素,制备了含有 LMN-1 的亲和层析柱,并使用质谱法分析了总蛋白提取物。
在孵育时,核糖体型 012 和 027 的菌株与葡萄糖补充剂(0.2%、0.5%和 1%)具有最佳的粘附性,而 RT135 的粘附性较差。RT014 和 RT133 不符合标准。在没有葡萄糖的情况下,没有任何核糖体型的粘附,这表明葡萄糖是必需的,并且在粘附中起重要作用。
这些发现表明,在葡萄糖存在的情况下,每种艰难梭菌核糖体型与 LMN-1 的相互作用方式不同,负责识别的粘附素可能是 SlpA 蛋白。