Pornwiroon Walairat, Bourchookarn Apichai, Paddock Christopher D, Macaluso Kevin R
Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 70803, USA .
Infect Immun. 2009 Dec;77(12):5262-71. doi: 10.1128/IAI.00911-09. Epub 2009 Sep 21.
Rickettsia parkeri, a recently recognized pathogen of human, is one of several Rickettsia spp. in the United States that causes a spotted fever rickettsiosis. To gain insights into its biology and pathogenesis, we applied the proteomics approach to establish a two-dimensional gel proteome reference map and combined this technique with cell surface biotinylation to identify surface-exposed proteins of a low-passage isolate of R. parkeri obtained from a patient. We identified 91 proteins by matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry. Of these, 28 were characterized as surface proteins, including virulence-related proteins (e.g., outer membrane protein A [OmpA], OmpB, beta-peptide, and RickA). Two-dimensional immunoblotting with serum from the R. parkeri-infected index patient was utilized to identify the immunoreactive proteins as potential targets for diagnosis and vaccine development. In addition to the known rickettsial antigens, OmpA and OmpB, we identified translation initiation factor 2, cell division protein FtsZ, and cysteinyl-tRNA synthetase as immunoreactive proteins. The proteome map with corresponding cell surface protein analysis and antigen detection will facilitate a better understanding of the mechanisms of rickettsial pathogenesis.
帕克立克次体是一种最近才被确认的人类病原体,是美国几种能引起斑点热立克次体病的立克次体之一。为深入了解其生物学特性和发病机制,我们应用蛋白质组学方法建立了二维凝胶蛋白质组参考图谱,并将该技术与细胞表面生物素化相结合,以鉴定从一名患者身上获得的低传代帕克立克次体分离株的表面暴露蛋白。我们通过基质辅助激光解吸电离串联飞行时间质谱鉴定了91种蛋白质。其中,28种被鉴定为表面蛋白,包括与毒力相关的蛋白(如外膜蛋白A [OmpA]、OmpB、β - 肽和RickA)。利用来自帕克立克次体感染的索引患者血清进行二维免疫印迹,以鉴定免疫反应性蛋白作为诊断和疫苗开发的潜在靶点。除了已知的立克次体抗原OmpA和OmpB外,我们还鉴定出翻译起始因子2、细胞分裂蛋白FtsZ和半胱氨酰 - tRNA合成酶为免疫反应性蛋白。带有相应细胞表面蛋白分析和抗原检测的蛋白质组图谱将有助于更好地理解立克次体发病机制。