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Green Tea Catechins Quench the Fluorescence of Bacteria-Conjugated Alexa Fluor Dyes

机译:绿茶儿茶素淬灭细菌结合的Alexa荧光染料的荧光

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摘要

Accumulating evidence suggests that Green tea polyphenolic catechins, especially the (-)-epigallocatechin gallate (EGCG), can be cross-linked to many proteins, and confer a wide range of anti-bacterial activities possibly by damaging microbial cytoplasmic lipids and proteins. At the doses that conferred protection against lethal polymicrobial infection (induced by cecal ligation and puncture), EGCG significantly reduced bacterial loads particularly in the liver and lung. To elucidate its bactericidal mechanisms, we determined whether EGCG affected the fluorescence intensities of bacteria-conjugated Alexa Fluor 488 or 594 dyes. When mixed with unconjugated Alexa Fluor 488 or 594 dyes, EGCG or analogs did not affect the fluorescence intensity of these dyes. In a sharp contrast, EGCG and some analogs (e.g., Catechin Gallate, CG), markedly reduced the fluorescence intensity of Gram-positive Staphylococcus aureus-conjugated Alexa 594 and Gram-negative Escherichia coli-conjugated Alexa 488. Interestingly, co-treatment with ethanol impaired the EGCG-mediated fluorescence quenching of the G+ S. aureus, but not of the G- E. coli-conjugated Alexa Flour dyes. In light of the notion that Alexa Fluor dyes can be quenched by aromatic amino acids, it is plausible that EGCG exerts anti-microbial activities possibly by altering microbial protein conformations and functions. This possibility can now be explored by screening other fluorescence-quenching agents for possible antimicrobial activities.
机译:越来越多的证据表明,绿茶多酚儿茶素,尤其是(-)-表没食子儿茶素没食子酸酯(EGCG)可以与许多蛋白质发生交联,并可能通过破坏微生物细胞质脂质和蛋白质而赋予广泛的抗菌活性。在给予防止致命性微生物感染(盲肠结扎和穿刺诱导)的剂量下,EGCG显着降低了细菌负荷,尤其是在肝脏和肺部。为了阐明其杀菌机理,我们确定了EGCG是否会影响细菌结合的Alexa Fluor 488或594染料的荧光强度。当与未结合的Alexa Fluor 488或594染料混合时,EGCG或类似物不会影响这些染料的荧光强度。与之形成鲜明对比的是,EGCG和某些类似物(例如Catechin Gallate,CG)显着降低了革兰氏阳性金黄色葡萄球菌结合的Alexa 594和革兰氏阴性大肠杆菌结合的Alexa 488的荧光强度。乙醇损害了EGCG介导的G + 金黄色葡萄球菌的荧光猝灭,但不损害G -大肠杆菌共轭的Alexa Flour染料的荧光猝灭。考虑到Alexa Fluor染料可以被芳香族氨基酸淬灭的观点,EGCG可能通过改变微生物蛋白的构象和功能发挥抗微生物活性。现在可以通过针对可能的抗菌活性筛选其他荧光猝灭剂来探索这种可能性。

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