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首页> 外文期刊>Infection and immunity >Changes in the surface of Leptospira interrogans serovar grippotyphosa during in vitro cultivation.
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Changes in the surface of Leptospira interrogans serovar grippotyphosa during in vitro cultivation.

机译：体外培养过程中问号钩端螺旋体血清型握轮虫表面的变化。

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Surface components of virulent and attenuated Leptospira interrogans serovar grippotyphosa were compared by using Triton X-114 solubilization and phase partitioning, immunoprecipitation of intact organisms, and freeze-fracture electron microscopy. Removal of the leptospiral outer membrane by using 0.1% Triton X-114 was demonstrated by whole-mount electron microscopy and by essentially complete solubilization of a lipopolysaccharidelike substance (LLS) from the outer membrane. Triton X-114 (0.1%) did not solubilize subsurface proteins, such as endoflagellar filaments or penicillin-binding proteins, which are markers for the periplasmic space and inner membrane, respectively. Triton X-114 solubilized material from both the virulent and attenuated strains, which partitioned into the hydrophobic, detergent phase, contained LLS and major proteins of 41 and 44 kDa, which were also immunoprecipitable from intact organisms. The virulent strain contained greater amounts of an LLS component with an apparent molecular mass of 30 kDa (R(f) = 0.57), whereas the attenuated strain contained larger amounts of an LLS component with an apparent molecular mass of 20 kDa (R(f) = 0.74). Differences in protein components between virulent and attenuated organisms were also detected; whereas the 41- and 44-kDa proteins were immunoprecipitated in equal amounts from both the virulent and attenuated strains, a 33-kDa protein was immunoprecipitated in significantly greater amounts from the attenuated strain. Quantitation of outer membrane particle density by freeze-fracture electron microscopy showed that both strains had a low transmembrane outer membrane protein content compared with that of typical gram-negative bacteria. The virulent and attenuated strains had 443 and 990 particles (P less than 0.000001) per micron, respectively, in the concave outer membrane fracture face. These findings suggest that in vitro cultivation of L. interrogans is accompanied by quantitative and qualitative changes in both LLS and outer membrane-associated proteins.
机译：通过使用Triton X-114增溶和相分配，完整生物体的免疫沉淀以及冷冻断裂电子显微镜，比较了有毒和减毒的问号钩端螺旋体血清型抓地力表层的表面成分。通过安装式电子显微镜和从外膜基本上完全溶解脂多糖样物质（LLS）证实了使用0.1％Triton X-114去除钩端螺旋体外膜。 Triton X-114（0.1％）不能溶解亚表面蛋白，例如内鞭毛丝或青霉素结合蛋白，它们分别是周质空间和内膜的标志。来自有毒和减毒菌株的Triton X-114增溶材料（分为疏水性去污剂相）包含LLS和41和44 kDa的主要蛋白质，它们也可以从完整的生物体中免疫沉淀。毒性菌株含有大量的表观分子量为30 kDa的LLS组分（R（f）= 0.57），而减毒菌株含有大量的表观分子量为20 kDa的LLS组分（R（f ）= 0.74）。还检测到有毒生物和减毒生物之间蛋白质成分的差异。而41 kDa和44 kDa蛋白则从毒性和减毒菌株中以等量免疫沉淀，而33 kDa蛋白则从减毒株中以明显更多的量进行免疫沉淀。通过冷冻断裂电子显微镜对外膜颗粒密度的定量显示，与典型的革兰氏阴性菌相比，这两种菌株的跨膜外膜蛋白含量均较低。在凹形外膜破裂面上，强毒株和减毒株分别具有每微米443和990个颗粒（P小于0.000001）。这些发现表明，在体外培养的问号乳酸菌伴随着LLS和外膜相关蛋白的定量和定性变化。

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