Definition of the extracellular proteome of pathogenic-phase Histoplasma capsulatum.
Definition of the extracellular proteome of pathogenic-phase Histoplasma capsulatum.
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DOI：
10.1021/pr1011697
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发表时间：
2011-04-01
影响因子：
4.4
通讯作者：
Rappleye, Chad A.
中科院分区：
文献类型：
--
作者：
Holbrook, Eric D.;Edwards, Jessica A.;Youseff, Brian H.;Rappleye, Chad A.
二型真菌病原体荚膜组织胞浆菌引起呼吸道和全身性疾病。在哺乳动物宿主内，致病性组织胞浆菌酵母感染宿主吞噬细胞、在其中复制并最终杀死宿主吞噬细胞。令人惊讶的是，很少有因素已被确定，有助于组织胞浆菌的毒力。为了解决这一缺陷，我们已经定义了细胞外蛋白质组的成分，使用LC-MS/MS分析组织胞浆菌致病期培养物中的蛋白质。致病性组织胞浆菌的胞外蛋白质组除分泌型Cbp 1外，还包括33个推导蛋白。这些蛋白质包括聚糖酶、与氧化应激防御相关的胞外酶、脱氢酶、伴侣样因子和五种新型培养滤液蛋白（Cfp）。对于蛋白质组学衍生的身份的独立验证，我们采用了基于RNA干扰（RNAi）的候选因子耗竭，并显示无细胞培养滤液中特定蛋白质的损失。定量RT-PCR显示，10个细胞外因子的表达是特别丰富的致病性酵母细胞相比，非致病性组织胞浆菌菌丝体，这表明这些蛋白质与组织胞浆菌的发病机制。此外，组织胞浆菌酵母表达这些因子在巨噬细胞内和感染小鼠肺。由于胞外蛋白位于宿主和病原体之间的界面，致病相胞外蛋白质组的定义为组织胞浆菌如何改变宿主-病原体相互作用以使其有利的分子解剖提供了基础。通过对组织胞浆菌酵母培养物的LC-MS/MS分析和对单个致病相蛋白条带的质谱分析，确定了致病相荚膜组织胞浆菌的胞外蛋白质组。组成蛋白质组的主要胞外蛋白包括聚糖酶、氧化应激蛋白和五种新蛋白。10种蛋白质在致病阶段优先表达，其中9种在感染过程中表达，表明它们有助于组织胞浆菌的发病机制。
The dimorphic fungal pathogen Histoplasma capsulatum causes respiratory and systemic disease. Within the mammalian host, pathogenic Histoplasma yeast infect, replicate within, and ultimately kill host phagocytes. Surprisingly, few factors have been identified that contribute to Histoplasma virulence. To address this deficiency, we have defined the constituents of the extracellular proteome using LC−MS/MS analysis of the proteins in pathogenic-phase culture filtrates of Histoplasma. In addition to secreted Cbp1, the extracellular proteome of pathogenic Histoplasma yeast consists of 33 deduced proteins. The proteins include glycanases, extracellular enzymes related to oxidative stress defense, dehydrogenase enzymes, chaperone-like factors, and five novel culture filtrate proteins (Cfp’s). For independent verification of proteomics-derived identities, we employed RNA interference (RNAi)-based depletion of candidate factors and showed loss of specific proteins from the cell-free culture filtrate. Quantitative RT-PCR revealed the expression of 10 of the extracellular factors was particularly enriched in pathogenic yeast cells as compared to nonpathogenic Histoplasma mycelia, suggesting that these proteins are linked to Histoplasma pathogenesis. In addition, Histoplasma yeast express these factors within macrophages and during infection of murine lungs. As extracellular proteins are positioned at the interface between host and pathogen, the definition of the pathogenic-phase extracellular proteome provides a foundation for the molecular dissection of how Histoplasma alters the host-pathogen interaction to its advantage. The extracellular proteome was defined for pathogenic-phase Histoplasma capsulatum through LC−MS/MS analysis of Histoplasma yeast culture filtrates and mass spectrometry of individual pathogenic-phase protein bands. The major extracellular proteins constituting the proteome include glycanases, oxidative stress proteins and five novel proteins. Ten proteins are preferentially expressed in the pathogenic phase of which nine are expressed during infection, suggesting that they contribute to Histoplasma pathogenesis.
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Dectin-1介导β-葡聚糖的生物学作用。
DOI：
10.1084/jem.20021890
发表时间：
2003-05-05
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Brown GD;Herre J;Williams DL;Willment JA;Marshall AS;Gordon S
通讯作者：
Gordon S
影响因子：
4.4
作者：
Da Silva, Carla A.;Chalouni, Cecile;Elias, Jack A.
通讯作者：
Elias, Jack A.
影响因子：
4.4
作者：
Crameri, R;Hemmann, S;Blaser, K
通讯作者：
Blaser, K
影响因子：
9.4
作者：
Chandrashekar, R;Curtis, KC;Weil, GJ
通讯作者：
Weil, GJ
影响因子：
3.1
作者：
Bohse, Megan L.;Woods, Jon P.
通讯作者：
Woods, Jon P.