Virta M, Akerman K E, Saviranta P, Oker-Blom C, Karp M T
Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland.
J Antimicrob Chemother. 1995 Aug;36(2):303-15. doi: 10.1093/jac/36.2.303.
A new method for studying the action of membranolytic agents by simple measurement of light emitted from cells is described. It is based on the expression of the click beetle (Pyrophorus plagiophthalamus) luciferase gene (lucGR) in Escherichia coli, Bacillus subtilis and Spodoptera frugiperda cells in order to make them bioluminescent. The diffusion of the substrate for luciferase enzyme through the cell membranes is very low at physiological pH, and therefore a change in membrane permeability is seen as a change of in-vivo luminescence of cells. The cells used in this study represent different membrane structures, and thus allow a comparison of the reactions of the different membranes towards membranolytic agents in a real-time measurement. The dose-response data correlated well with target cell viable count. In addition, the time course of light emission as a consequence of permeabilizing compound is dose-dependent. The action of the compounds on prokaryotic and eukaryotic cells was found to be highly dependent on the permeabilizer used.
本文描述了一种通过简单测量细胞发出的光来研究膜溶解剂作用的新方法。该方法基于将叩头虫（Pyrophorus plagiophthalamus）荧光素酶基因（lucGR）在大肠杆菌、枯草芽孢杆菌和草地贪夜蛾细胞中表达，使其具有生物发光特性。在生理pH值下，荧光素酶底物透过细胞膜的扩散非常低，因此膜通透性的变化表现为细胞体内发光的变化。本研究中使用的细胞代表不同的膜结构，从而能够在实时测量中比较不同膜对膜溶解剂的反应。剂量-反应数据与靶细胞存活计数相关性良好。此外，由于通透化合物导致的发光时间进程呈剂量依赖性。发现这些化合物对原核细胞和真核细胞的作用高度依赖于所使用的通透剂。
