Brissette Catherine A, Haupt Katrin, Barthel Diana, Cooley Anne E, Bowman Amy, Skerka Christina, Wallich Reinhard, Zipfel Peter F, Kraiczy Peter, Stevenson Brian
Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky College of Medicine, MN 469, W. R. Willard Medical Education Building, Lexington, KY 40536-0298, USA.
Infect Immun. 2009 Jan;77(1):300-6. doi: 10.1128/IAI.01133-08. Epub 2008 Nov 10.
Host-derived plasmin plays a critical role in mammalian infection by Borrelia burgdorferi. The Lyme disease spirochete expresses several plasminogen-binding proteins. Bound plasminogen is converted to the serine protease plasmin and thereby may facilitate the bacterium's dissemination throughout the host by degrading extracellular matrix. In this work, we demonstrate plasminogen binding by three highly similar borrelial outer surface proteins, ErpP, ErpA, and ErpC, all of which are expressed during mammalian infection. Extensive characterization of ErpP demonstrated that this protein bound in a dose-dependent manner to lysine binding site I of plasminogen. Removal of three lysine residues from the carboxy terminus of ErpP significantly reduced binding of plasminogen, and the presence of a lysine analog, epsilon-aminocaproic acid, inhibited the ErpP-plasminogen interaction, thus strongly pointing to a primary role for lysine residues in plasminogen binding. Ionic interactions are not required in ErpP binding of plasminogen, as addition of excess NaCl or the polyanion heparin did not have any significant effect on binding. Plasminogen bound to ErpP could be converted to the active enzyme, plasmin. The three plasminogen-binding Erp proteins can also bind the host complement regulator factor H. Plasminogen and factor H bound simultaneously and did not compete for binding to ErpP, indicating separate binding sites for both host ligands and the ability of the borrelial surface proteins to bind both host proteins.
宿主来源的纤溶酶在哺乳动物感染伯氏疏螺旋体过程中发挥关键作用。莱姆病螺旋体表达多种纤溶酶原结合蛋白。结合的纤溶酶原被转化为丝氨酸蛋白酶纤溶酶,从而可能通过降解细胞外基质促进细菌在宿主体内传播。在这项研究中,我们证明了三种高度相似的疏螺旋体外膜蛋白ErpP、ErpA和ErpC能结合纤溶酶原,这三种蛋白在哺乳动物感染期间均有表达。对ErpP的广泛表征表明,该蛋白以剂量依赖的方式结合到纤溶酶原的赖氨酸结合位点I。从ErpP的羧基末端去除三个赖氨酸残基显著降低了纤溶酶原的结合,赖氨酸类似物ε-氨基己酸的存在抑制了ErpP与纤溶酶原的相互作用,因此有力地表明赖氨酸残基在纤溶酶原结合中起主要作用。ErpP与纤溶酶原结合不需要离子相互作用,因为添加过量的NaCl或多聚阴离子肝素对结合没有任何显著影响。结合到ErpP上的纤溶酶原可以转化为活性酶纤溶酶。三种结合纤溶酶原的Erp蛋白也能结合宿主补体调节因子H。纤溶酶原和因子H同时结合,且不竞争与ErpP的结合,这表明宿主配体有单独的结合位点,且疏螺旋体表面蛋白有结合两种宿主蛋白的能力。