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Rhodamine 6G

罗丹明6G

Company: Sigma-Aldrich
Catalog#: R4127
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Measurement of Energy-dependent Rhodamine 6G Efflux in Yeast Species
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Date:
2017-08-05
[Abstract]  Rhodamine 6G is a highly fluorescent dye often used to determine the transport activity of yeast membrane efflux pumps. The ATP-binding cassette transporter KlPdr5p confers resistance to several unrelated drugs in Kluyveromyces lactis. KlPdr5p also extrudes rhodamine 6G (R6G) from intact yeast cells in an energy-dependent manner. Incubation of yeast cells in the presence of 2-deoxy-D-glucose (inhibitor of glycolysis) and R6G (mitochondrial ATPase inhibitor) leads to marked depletion of intracellular ATP pool (Kolaczkowski et al., 1996). An active KlPdr5p mediated extrusion of R6G from intact yeast cells can be followed by direct measurement of the fluorescence of extruded R6G in the assay buffer. [摘要]  罗丹明6G是一种高荧光染料,常用于测定酵母膜外排泵的运输活性。 ATP结合盒转运蛋白Klr5p赋予乳酸克鲁维酵酵菌中几种不相关药物的抗性。 Pdr5p还以能量依赖的方式从完整的酵母细胞中挤出罗丹明6G(R6G)。 在2-脱氧-D-葡萄糖(糖酵解抑制剂)和R6G(线粒体ATP酶抑制剂)存在下酵母细胞的孵育导致细胞内ATP池的显着消耗(Kolaczkowski等人,1996)。 从完整的酵母细胞中可以直接测量R6G的荧光的直接测量PdF5p介导的来自完整酵母细胞的R6G的挤出。
【背景】多药物外排泵广泛分布,可在所有生物种类中找到。它们代表抗菌素耐药性的重要机制。量化外排泵活性的能力对于理解其对生理过程的贡献和评估潜在治疗药物(例如,外排抑制剂)的有效性(Blair和Piddock,2016)是必要的。外排活动测量的方法主要依赖于两种不同的机制。一些方法直接测量底物流出,即,多少基质被泵出,其他方法测量细胞内的底物分子积累,其水平然后用于间接推断流出。然而,后者由于可改变膜渗透性而改变了染料流入速率(Blair和Piddock,2016)的敏感性较差。 R6G在生长中的积累。白念珠菌细胞与ABC转运蛋白念珠菌药物抗性1(CDR1 ...

Extraction and Purification of Mycobacterial Mycolic Acids
Author:
Date:
2014-10-20
[Abstract]  Mycolic acids are major long-chain fatty acids, containing up to 80-90 carbon atoms that represent essential components of the mycobacterial cell wall (Pawelczyk and Kremer, 2014). Each mycobacterial species possesses a specific mycolic acid profile characterized by various chemical modifications that decorate the lipid. Mycolic acids play a critical role in the architecture and impermeability of the cell envelope, hence the natural resistance of mycobacteria to most antibiotic treatments. They are also key determinants of virulence in pathogenic species, including Mycobacterium tuberculosis (M. tuberculosis), the causative agent of tuberculosis. In addition, they are known as the primary target of several first-line and second-line antitubercular drugs. Thus, the unique ... [摘要]  霉菌酸是主要的长链脂肪酸,含有高达80-90个碳原子,代表分枝杆菌细胞壁的基本组分(Pawelczyk和Kremer,2014)。每种分枝杆菌物种具有特征性霉菌酸分布,其特征在于装饰脂质的各种化学修饰。霉菌酸在细胞包膜的结构和不可渗透性中起关键作用,因此分枝杆菌对大多数抗生素治疗的天然抗性。它们也是在致病物种中的毒力的关键决定因素,包括结核分枝杆菌(结核分枝杆菌),结核病的致病因子。此外,它们被认为是几种一线和二线抗结核药物的主要靶点。因此,参与霉菌酸生物合成途径的独特的酶代表了用于未来化学疗法的靶的有吸引力的储库,其发展在M的多药耐药和广泛耐药菌株的背景下是特别保证的。肺结核。在这里,我们描述了从分枝杆菌中提取霉菌酸的方案。各种亚种的纯化对于涉及质谱或NMR的后续结构研究可能是特别有用的。通过薄层色谱的霉菌酸模式的定性和定量生物化学表征可用于解决药物如何改变霉菌酸生物合成(Alahari等人,2007,Hartkoorn等人, (Bhatt等人,2007)或解开新的霉菌酸调节机制(Vilcheze等人,2012),以研究在该代谢途径中受影响的遗传修饰的突变体的表型(Bhatt等人, ,2014)。相同的方案可以应用于所有分枝杆菌,包括环境和致病物种。

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