Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada.
PLoS One. 2013 Jul 22;8(7):e67190. doi: 10.1371/journal.pone.0067190. Print 2013.
PfbA (plasmin- and fibronectin-binding protein A) is an extracellular Streptococcus pneumoniae cell-wall attached surface protein that binds to fibronectin, plasmin, and plasminogen. Here we present a structural analysis of the surface exposed domains of PfbA using a combined approach of X-ray crystallography and small-angle X-ray scattering (SAXS). The crystal structure of the PfbA core domain, here called PfbAβ, determined to 2.28 Å resolution revealed an elongated 12-stranded parallel β-helix fold, which structure-based comparisons reveal is most similar to proteins with carbohydrate modifying activity. A notable feature of the PfbAβ is an extensive cleft on one face of the protein with electrochemical and spatial features that are analogous to structurally similar carbohydrate-active enzymes utilizing this feature for substrate accommodation. Though this cleft displays a combination of basic amino acid residues and solvent exposed aromatic amino acids that are distinct features for recognition of carbohydrates, no obvious arrangement of amino acid side chains that would constitute catalytic machinery is evident. The pseudo-atomic SAXS model of a larger fragment of PfbA suggests that it has a relatively well-ordered structure with the N-terminal and core domains of PfbA adopting an extend organization and reveals a novel structural class of surface exposed pneumococcal matrix molecule adhesins.
PfbA(纤溶酶和纤维结合蛋白结合蛋白 A)是一种附着在肺炎链球菌细胞壁上的细胞外表面蛋白,可与纤维结合蛋白、纤溶酶和纤溶酶原结合。在这里,我们使用 X 射线晶体学和小角 X 射线散射(SAXS)的组合方法,对 PfbA 的表面暴露结构域进行了结构分析。PfbA 核心结构域(称为 PfbAβ)的晶体结构,分辨率为 2.28Å,揭示了一个长的 12 股平行β-螺旋折叠,基于结构的比较表明,它与具有碳水化合物修饰活性的蛋白质最为相似。PfbAβ 的一个显著特征是在蛋白质的一个面上有一个广泛的裂缝,其电化学和空间特征与利用该特征容纳底物的结构相似的碳水化合物活性酶相似。尽管这个裂缝显示出碱性氨基酸残基和溶剂暴露的芳香族氨基酸的组合,这些是识别碳水化合物的明显特征,但没有明显的催化机制的氨基酸侧链排列是明显的。较大片段的 PfbA 的拟原子 SAXS 模型表明,它具有相对有序的结构,PfbA 的 N 端和核心结构域采用伸展组织,并揭示了一种新型的表面暴露肺炎球菌基质分子粘附素的结构类别。