Identification and characterization of a cell surface protein of Prevotella intermedia 17 with broad-spectrum binding activity for extracellular matrix proteins.
Identification and characterization of a cell surface protein of Prevotella intermedia 17 with broad-spectrum binding activity for extracellular matrix proteins.
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具有广谱细胞外基质蛋白结合活性的中间普雷沃氏菌 17 的细胞表面蛋白的鉴定和表征。
DOI：
10.1002/pmic.200600177
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发表时间：
2006
期刊：
影响因子：
3.4
通讯作者：
Lewis,JaninaP
中科院分区：
文献类型：
--
作者：
Yu,Fan;Iyer,Divya;Anaya,Cecilia;Lewis,JaninaP
中间普雷沃氏菌结合并侵入多种宿主细胞。这种结合很可能是通过称为粘附素的细胞表面蛋白介导的。为了鉴定与宿主细胞外基质 （ECM）成分纤连蛋白结合的蛋白质，并研究细菌定植的分子机制，我们应用蛋白质组学方法对P. 中间菌株 17 外膜蛋白。 2-DE 随后使用纤连蛋白作为探针进行 Far Western 印迹分析，揭示了 29-kDa 纤连蛋白结合蛋白，此处指定为 AdpB。使用 PMF 确定蛋白质的分子身份，然后搜索 P。 intermedia17 蛋白质数据库。数据库搜索揭示了 AdpB 与多种细菌外膜蛋白的相似性，包括空肠弯曲杆菌的纤连蛋白结合蛋白。重组 AdpB 蛋白以可饱和、剂量依赖的方式结合纤连蛋白以及其他宿主 ECM 成分，包括纤维蛋白原和层粘连蛋白。 AdpB 与固定化纤连蛋白的结合也受到可溶性纤连蛋白、层粘连蛋白和纤维蛋白原的抑制，表明这种结合是特异性的。最后，使用抗 AdpB 的免疫电子显微镜证实了该蛋白质的细胞表面位置。这是第一个在 P 中鉴定和表征的具有广谱 ECM 结合能力的细胞表面蛋白。 中介17.
Prevotella intermediabinds and invades a variety of host cells. This binding is most probably mediated through cell surface proteins termed adhesins. To identify proteins binding to the host extracellular matrix (ECM) component, fibronectin, and study the molecular mechanism underlying bacterial colonization, we applied proteomic approaches to perform a global investigation ofP. intermediastrain 17 outer membrane proteins. 2‐DE followed by Far Western Blot analysis using fibronectin as a probe revealed a 29‐kDa fibronectin‐binding protein, designated here AdpB. The molecular identity of the protein was determined using PMF followed by a search of theP. intermedia17 protein database. Database searches revealed the similarity of AdpB to multiple bacterial outer membrane proteins including the fibronectin‐binding protein fromCampylobacter jejuni. A recombinant AdpB protein bound fibronectin as well as other host ECM components, including fibrinogen and laminin, in a saturable, dose‐dependent manner. Binding of AdpB to immobilized fibronectin was also inhibited by soluble fibronectin, laminin, and fibrinogen, indicating the binding was specific. Finally, immunoelectron microscopy with anti‐AdpB demonstrated the cell surface location of the protein. This is the first cell surface protein with a broad‐spectrum ECM‐binding abilities identified and characterized inP. intermedia17.