Back Catherine R, Sztukowska Maryta N, Till Marisa, Lamont Richard J, Jenkinson Howard F, Nobbs Angela H, Race Paul R
From the School of Oral and Dental Sciences, University of Bristol, Lower Maudlin Street, Bristol BS1 2LY, United Kingdom.
the Department of Oral Immunology and Infectious Diseases, University of Louisville, Louisville, Kentucky 40202; the Department of Dentistry, University of Information Technology and Management, 35-225 Rzeszow, Poland.
J Biol Chem. 2017 Feb 3;292(5):1538-1549. doi: 10.1074/jbc.M116.760975. Epub 2016 Dec 5.
Adherence of bacteria to biotic or abiotic surfaces is a prerequisite for host colonization and represents an important step in microbial pathogenicity. This attachment is facilitated by bacterial adhesins at the cell surface. Because of their size and often elaborate multidomain architectures, these polypeptides represent challenging targets for detailed structural and functional characterization. The multifunctional fibrillar adhesin CshA, which mediates binding to both host molecules and other microorganisms, is an important determinant of colonization by Streptococcus gordonii, an oral commensal and opportunistic pathogen of animals and humans. CshA binds the high-molecular-weight glycoprotein fibronectin (Fn) via an N-terminal non-repetitive region, and this protein-protein interaction has been proposed to promote S. gordonii colonization at multiple sites within the host. However, the molecular details of how these two proteins interact have yet to be established. Here we present a structural description of the Fn binding N-terminal region of CshA, derived from a combination of X-ray crystallography, small angle X-ray scattering, and complementary biophysical methods. In vitro binding studies support a previously unreported two-state "catch-clamp" mechanism of Fn binding by CshA, in which the disordered N-terminal domain of CshA acts to "catch" Fn, via formation of a rapidly assembled but also readily dissociable pre-complex, enabling its neighboring ligand binding domain to tightly clamp the two polypeptides together. This study presents a new paradigm for target binding by a bacterial adhesin, the identification of which will inform future efforts toward the development of anti-adhesive agents that target S. gordonii and related streptococci.
细菌粘附于生物或非生物表面是其在宿主体内定殖的前提条件,也是微生物致病过程中的重要一步。这种粘附作用由细胞表面的细菌粘附素介导。由于这些多肽的大小以及通常复杂的多结构域结构,它们成为详细进行结构和功能表征的具有挑战性的目标。多功能纤维状粘附素CshA介导与宿主分子和其他微生物的结合,是动物和人类口腔共生菌及机会致病菌戈登氏链球菌定殖的重要决定因素。CshA通过N端非重复区域结合高分子量糖蛋白纤连蛋白(Fn),并且这种蛋白质 - 蛋白质相互作用被认为可促进戈登氏链球菌在宿主体内多个部位的定殖。然而,这两种蛋白质相互作用的分子细节尚未确定。在此,我们结合X射线晶体学、小角X射线散射和互补生物物理方法,对CshA的Fn结合N端区域进行了结构描述。体外结合研究支持了一种此前未报道的CshA结合Fn的双态“捕获 - 钳夹”机制,其中CshA无序的N端结构域通过形成快速组装但也易于解离的预复合物来“捕获”Fn,使其相邻的配体结合结构域能够将这两种多肽紧密钳夹在一起。这项研究提出了细菌粘附素与靶标结合的新范式,其鉴定将为未来开发针对戈登氏链球菌及相关链球菌的抗粘附剂的努力提供信息。