Institute of Medical Microbiology, Jena University Hospital, Jena, Germany.
Center for Infectious Diseases and Infection Control, Jena University Hospital, Jena, Germany.
Sci Rep. 2017 Oct 20;7(1):13665. doi: 10.1038/s41598-017-14168-4.
The extracellular matrix protein Emp of Staphylococcus aureus is a secreted adhesin that mediates interactions between the bacterial surface and extracellular host structures. However, its structure and role in staphylococcal pathogenesis remain unknown. Using multidisciplinary approaches, including circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy, transmission electron (TEM) and immunogold transmission electron microscopy, functional ELISA assays and in silico techniques, we characterized the Emp protein. We demonstrated that Emp and its truncated forms bind to suprastructures in human skin, cartilage or bone, among which binding activity seems to be higher for skin compounds. The binding domain is located in the C-terminal part of the protein. CD spectroscopy revealed high contents of β-sheets (39.58%) and natively disordered structures (41.2%), and TEM suggested a fibrous structure consisting of Emp polymers. The N-terminus seems to be essential for polymerization. Due to the uncommonly high histidine content, we suggest that Emp represents a novel type of histidine-rich protein sharing structural similarities to leucine-rich repeats proteins as predicted by the I-TASSER algorithm. These new findings suggest a role of Emp in infections of deeper tissue and open new possibilities for the development of novel therapeutic strategies.
金黄色葡萄球菌的细胞外基质蛋白 Emp 是一种分泌型黏附素,介导细菌表面与细胞外宿主结构之间的相互作用。然而,其结构及其在金黄色葡萄球菌发病机制中的作用仍不清楚。我们采用多种方法,包括圆二色性(CD)和傅里叶变换红外(FTIR)光谱、透射电子(TEM)和免疫金透射电子显微镜、功能性 ELISA 测定和计算技术,对 Emp 蛋白进行了表征。我们证明 Emp 及其截断形式与人体皮肤、软骨或骨骼中的超结构结合,其中与皮肤化合物的结合活性似乎更高。结合域位于蛋白质的 C 端部分。CD 光谱显示β-折叠(39.58%)和天然无规结构(41.2%)含量高,TEM 表明存在由 Emp 聚合物组成的纤维结构。N 端似乎对聚合至关重要。由于组氨酸含量异常高,我们推测 Emp 代表了一种新型的富含组氨酸的蛋白质,与 I-TASSER 算法预测的富含亮氨酸重复蛋白具有结构相似性。这些新发现表明 Emp 在深部组织感染中的作用,并为开发新型治疗策略开辟了新的可能性。