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蛋白组学研究表明嗜肺军团菌感染棘阿米巴属的过程

Proteomic aspects of Parachlamydia acanthamoebae infection in Acanthamoeba spp.

机构信息

Department of Specific Prophylaxis and Tropical Medicine at the Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Kinderspitalgasse 15, Vienna A-1090, Austria.

出版信息

ISME J. 2010 Nov;4(11):1366-74. doi: 10.1038/ismej.2010.68. Epub 2010 May 20.

Abstract

The free-living but facultatively pathogenic amoebae of the genus Acanthamoeba are frequently infected with bacterial endosymbionts that can have a profound influence on the physiology and viability of their host. Parachlamydia acanthamoebae, a chlamydial endosymbiont in acanthamoebae, is known to be either symbiotic or lytic to its host, depending on the ambient conditions, for example, temperature. Moreover, parachlamydiae can also inhibit the encystment process in Acanthamoeba, an essential survival strategy of their host for the evasion of chemotherapeutic agents, heat, desiccation and radiation. To obtain a more detailed picture of the intracellular interactions of parachlamydiae and acanthamoebae, we studied parachlamydial infection in several Acanthamoeba isolates at the proteomic level by means of two-dimensional gel electrophoresis (2DE) and mass spectrometry. We observed that P. acanthamoebae can infect all three morphological subtypes of the genus Acanthamoeba and that the proteome pattern of released P. acanthamoebae elementary bodies was always practically identical regardless of the Acanthamoeba strain infected. Moreover, by comparing proteome patterns of encysting cells from infected and uninfected Acanthamoeba cultures, it was shown that encystment is blocked by P. acanthamoebae at a very early stage. Finally, on 2D-gels of purified P. acanthamoebae from culture supernatants, a subunit of the NADH-ubiquinone oxidoreductase complex, that is, an enzyme that has been described as an indicator for bacterial virulence was identified by a mass spectrometric and bioinformatic approach.

摘要

自由生活但兼性致病的棘阿米巴属阿米巴常被细菌内共生体感染,这些共生体对宿主的生理和生存能力有深远的影响。棘阿米巴中的共生衣原体 Parachlamydia acanthamoebae 对其宿主的影响可能是共生的,也可能是裂解的,这取决于环境条件,例如温度。此外,共生衣原体还可以抑制棘阿米巴的包囊形成过程,这是宿主逃避化学治疗剂、热、干燥和辐射等生存策略的重要手段。为了更详细地了解共生衣原体和棘阿米巴之间的细胞内相互作用,我们在蛋白质组学水平上通过二维凝胶电泳(2DE)和质谱法研究了几种棘阿米巴分离株中的共生衣原体感染。我们观察到 P. acanthamoebae 可以感染棘阿米巴属的所有三种形态亚型,并且释放的 P. acanthamoebae 原始体的蛋白质组图谱始终几乎完全相同,无论感染的棘阿米巴株如何。此外,通过比较感染和未感染的棘阿米巴培养物中包囊细胞的蛋白质组图谱,表明共生衣原体在非常早期就阻断了包囊形成。最后,在来自培养上清液的纯化 P. acanthamoebae 的 2D 凝胶上,通过质谱和生物信息学方法鉴定出 NADH-泛醌氧化还原酶复合物的一个亚基,即一种被描述为细菌毒力指标的酶。

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