Fluorescence-detected circular dichroism of ethidium in vivo and bound to deoxyribonucleic acid in vitro.
Fluorescence-detected circular dichroism of ethidium in vivo and bound to deoxyribonucleic acid in vitro.
复制标题
荧光检测体内乙锭的圆二色性以及体外与脱氧核糖核酸的结合。
DOI：
10.1021/bi00333a001
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发表时间：
1985
期刊：
影响因子：
2.9
通讯作者：
Turner,DH
中科院分区：
文献类型：
--
作者：
Lamos,ML;Turner,DH
Department of Chemistry, University of Rochester, Rochester, New York 14627 Received February 5, 1985 abstract: Fluorescence-detected circular dichroism (FDCD) spectra are reported for ethidium in Escherichia coli cells and bound to E. coli DNA in vitro. FDCD bands are observed at 325 and 385 nm. These bands change amplitude as the ethidium to DNA ratio changes. Spectra are similar for invivo and in vitro measurements. However, the bands at 325 and 385 nm disappear when ethidium binds to macromolecules without intercalating between base pairs. The results demonstrate thatFDCD spectra can be measured in cell suspensions and indicate that ethidium bindsto nucleic acids in E. coli cells by intercalation. i^ Tiysical-chemical studies have provided detailed information on the binding of drugs to nucleic acids in solution and in the solid state (Waring, 1981; Berman & Young, 1981; Krugh & Reinhardt, 1975; Jain et al., 1977; Tsai et al., 1977; Neidle et al., 1977; Wang et al., 1978; Quigley et al., 1980; Reinhardt & Krugh, 1978; Reuben et al., 1978; Pardi et al., 1983; Patel et al., 1981). These studies indicate drugs often bind to DNA by intercalation. The assumption is usually made that binding in vivo is similar. This is difficult to prove, especially since many additional receptors are present in a cell (Tritton & Yee, 1982). Fluorescence-detected circular dichroism (Turner et al., 1974; Turner, 1978)(FDCD) 1* is a spectroscopy that can be used both in vitro and in vivo and is sensitive to conformation (Turner et al., 1975; Lobenstine et al., 1981). This paper reports FDCD studies of ethidium bound to DNA in vitro and taken up by Escherichiacoli cells in vivo. It provides the first measurements of FDCD on an in vivo system. The results indicateethidium binds by intercalation in vivo as well as in vitro and demonstratethat FDCD can provide a spec-troscopic bridge between in vitro and in vivo studies. Materials and Methods
