IBET, Apartado 12, 2780-901 Oeiras, Portugal.
Vet Microbiol. 2012 May 4;156(3-4):305-14. doi: 10.1016/j.vetmic.2011.11.022. Epub 2011 Dec 1.
The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electrophoresis combined with mass spectrometry. Among the sixty-four spots detected, we could identify only four proteins from the MAP1-family; the other proteins detected were mainly related to energy, amino acid and general metabolism (26%), to protein turnover, chaperones and survival (21%) and to information processes (14%) or classified as hypothetical proteins (23%). Additional studies on MAP1-family protein using immunochemical labeling also revealed that these proteins are differentially expressed along the bacterium life cycle, presenting different structural organization. Interestingly, when infectious elementary bodies (EBs) are released from host cells, MAP1 appears to be organized in SDS and heat-resistant dimers and trimers stabilized by disulfide bridges. Overall, the results presented herein not only reveal the first partial proteome map of ER but provide new insights on the expression ER MAP1-family proteins in host endothelial cells.
立克次体目埃立克体属(ER)是心水病的病原体,心水病是撒哈拉以南非洲和加勒比地区牲畜的一种致命蜱传疾病,对畜牧业生产造成了严重的经济限制。为了鉴定该细菌表达的最主要蛋白,尤其是主要抗原蛋白 1(map1)多基因家族编码的蛋白,我们采用二维凝胶电泳结合质谱法构建了在血管内皮细胞中培养的 ER 的蛋白质组图谱。在检测到的 64 个斑点中,我们只能从 MAP1 家族中鉴定出 4 种蛋白;其他检测到的蛋白主要与能量、氨基酸和一般代谢(26%)、蛋白质周转、伴侣和生存(21%)以及信息处理(14%)有关,或归类为假定蛋白(23%)。使用免疫化学标记对 MAP1 家族蛋白的进一步研究还表明,这些蛋白在细菌生命周期中表达不同,呈现不同的结构组织。有趣的是,当传染性的始体(EBs)从宿主细胞中释放出来时,MAP1 似乎以二硫键稳定的 SDS 和耐热二聚体和三聚体的形式存在。总之,本文的结果不仅揭示了 ER 的第一张部分蛋白质组图谱,还为 ER MAP1 家族蛋白在宿主内皮细胞中的表达提供了新的见解。