Following Drug Uptake and Reactions inside Escherichia coli Cells by Raman Microspectroscopy
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- 作者：Hossein Heidari Torkabadi ; Christopher R. Bethel ; Krisztina M. Papp-Wallace ; Piet A. J. de Boer ; Robert A. Bonomo ; Paul R. Carey
- 刊名：Biochemistry
- 出版年：2014
- 出版时间：July 1, 2014
- 年：2014
- 卷：53
- 期：25
- 页码：4113-4121
- 全文大小：547K
- ISSN：1520-4995
文摘
Raman microspectroscopy combined with Raman difference spectroscopy reveals the details of chemical reactions within bacterial cells. The method provides direct quantitative data on penetration of druglike molecules into Escherichia coli cells in situ along with the details of drug鈥搕arget reactions. With this label-free technique, clavulanic acid and tazobactam can be observed as they penetrate into E. coli cells and subsequently inhibit 尾-lactamase enzymes produced within these cells. When E. coli cells contain a 尾-lactamase that forms a stable complex with an inhibitor, the Raman signature of the known enamine acyl鈥揺nzyme complex is detected. From Raman intensities it is facile to measure semiquantitatively the number of clavulanic acid molecules taken up by the lactamase-free cells during growth.