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Characterization of a Cytotoxic Factor in Culture Filtrates of Serratia marcescens

机译：粘质沙雷氏菌培养滤液中细胞毒性因子的表征

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Serratia marcescens culture filtrates have been reported to be cytotoxic to mammalian cells. Using biochemical and genetic approaches, we have identified a major source of this cytotoxic activity. Both heat and protease treatments abrogated the cytotoxicity of S. marcescens culture filtrates towards HeLa cells, suggesting the involvement of one or more protein factors. A screen for in vitro cytotoxic activity revealed that S. marcescens mutant strains that are deficient in production of a 56-kDa metalloprotease are significantly less cytotoxic to mammalian cells. Cytotoxicity was significantly reduced when culture filtrates prepared from wild-type strains were pretreated with either EDTA or 1,10-phenanthroline, which are potent inhibitors of the 56-kDa metalloprotease. Furthermore, cytotoxic activity was restored when the same culture filtrates were incubated with zinc divalent cations, which are essential for enzymatic activity of the 56-kDa metalloprotease. Finally, recombinant expression of the S. marcescens 56-kDa metalloprotease conferred a cytotoxic phenotype on the culture filtrates of a nonpathogenic Escherichia coli strain. Collectively, these data suggest that the 56-kDa metalloprotease contributes significantly to the in vitro cytotoxic activity commonly observed in S. marcescens culture filtrates.
机译：据报道，粘质沙雷氏菌培养滤液对哺乳动物细胞具有细胞毒性。使用生化和遗传方法，我们已经确定了这种细胞毒活性的主要来源。热处理和蛋白酶处理均消除了 S的细胞毒性。 marcescens 培养物向HeLa细胞滤出，提示一种或多种蛋白质因子参与其中。体外细胞毒性活性的筛选显示了 S。缺乏生产56 kDa金属蛋白酶的marcescens 突变株对哺乳动物细胞的细胞毒性明显降低。用EDTA或1,10-菲咯啉（它们是56 kDa金属蛋白酶的有效抑制剂）预处理从野生型菌株制备的培养滤液时，细胞毒性显着降低。此外，当将相同的培养滤液与二价锌阳离子温育时，恢复了细胞毒性活性，这对于56 kDa金属蛋白酶的酶活性至关重要。最后， S的重组表达。 marcescens 56 kDa的金属蛋白酶在非致病性大肠杆菌菌株的培养滤液中具有细胞毒性表型。总的来说，这些数据表明56kDa金属蛋白酶对 S中通常观察到的体外细胞毒性活性有显着贡献。粘菌丝培养物滤液。

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