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首页> 外文期刊>Infection and immunity >Aspergillus fumigatus complement inhibitor: production, characterization, and purification by hydrophobic interaction and thin-layer chromatography.
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Aspergillus fumigatus complement inhibitor: production, characterization, and purification by hydrophobic interaction and thin-layer chromatography.

机译：烟曲霉补体抑制剂：通过疏水相互作用和薄层色谱法进行生产，表征和纯化。

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Aspergillus fumigatus has previously been shown to produce a soluble extracellular inhibitor of the alternative complement pathway, called Aspergillus complement inhibitor, or CI. We now report an efficient method for production of CI which relies on the fact that poorly conidiating cultures yielded CI activity with approximately sevenfold-higher potency than CI produced by conidiating cultures. CI from poorly conidiating cultures provided 50% inhibition of alternative pathway-mediated binding of 125I-labeled complement component C3 to cryptococcal blastoconidia at a mean concentration of 60 micrograms/ml. The ability of crude CI to inhibit the alternative complement pathway seemed to be independent of intact protein or polysaccharide structure, as evidenced by resistance of inhibitory activity to digestion by proteases, including subtilisin, alpha-chymotrypsin, papain, and pepsin as well as endoglycosidases F and H. Separation of the active inhibitory component of CI from contaminating materials contained in crude CI preparations was achieved by using Phenylsuperose hydrophobic interaction chromatography in a fast protein liquid chromatography system. The active material proved to be extremely hydrophobic, desorbing from the column only during elution with ethanol; it contained only 15% protein and 5% polysaccharide. Furthermore, results from preparative thin-layer chromatography indicated that lipids which comigrated with phosphatidylserine/phosphatidylinositol and phosphatidylethanolamine possessed significant complement-inhibitory activity. Taken together, these data suggested that phospholipids from A. fumigatus contributed to the functional activity of CI.
机译：以前已证明烟曲霉会产生另一种补体途径的可溶性细胞外抑制剂，称为曲霉补体抑制剂或CI。现在，我们报告一种有效的CI生产方法，该方法依赖于这样的事实，即弱连接的培养物产生的CI活性比其伴随的培养物产生的CI活性高大约七倍。来自条件差的培养物的CI以60微克/毫升的平均浓度提供了50％的替代途径介导的125I标记的补体成分C3与隐球菌瘟病的结合。粗制CI抑制替代补体途径的能力似乎与完整的蛋白质或多糖结构无关，如蛋白酶对蛋白酶（包括枯草杆菌蛋白酶，α-胰凝乳蛋白酶，木瓜蛋白酶和胃蛋白酶以及内切糖苷酶F）的消化抑制活性的抵抗力所证明的那样。通过在快速蛋白质液相色谱系统中使用苯基超级糖疏水相互作用色谱法，可将CI的活性抑制成分与粗制CI制剂中所含的污染物质分离。活性物质被证明具有极强的疏水性，仅在用乙醇洗脱时才从色谱柱上解吸。它仅包含15％的蛋白质和5％的多糖。此外，制备薄层色谱的结果表明，与磷脂酰丝氨酸/磷脂酰肌醇和磷脂酰乙醇胺迁移的脂质具有明显的补体抑制活性。综上所述，这些数据表明来自烟曲霉的磷脂有助于CI的功能活性。

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