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Biochemistry. 2014 Jul 1;53(25):4113-21. doi: 10.1021/bi500529c. Epub 2014 Jun 18.

Following drug uptake and reactions inside Escherichia coli cells by Raman microspectroscopy.

Author information

1
Departments of Chemistry, §Molecular Biology and Microbiology, ∥Pharmacology, ⊥Medicine, and #Biochemistry, Case Western Reserve University , Cleveland, Ohio 44106, United States.

Abstract

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-target reactions. With this label-free technique, clavulanic acid and tazobactam can be observed as they penetrate into E. coli cells and subsequently inhibit β-lactamase enzymes produced within these cells. When E. coli cells contain a β-lactamase that forms a stable complex with an inhibitor, the Raman signature of the known enamine acyl-enzyme 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.

PMID:
24901294
PMCID:
PMC4082380
DOI:
10.1021/bi500529c
[Indexed for MEDLINE]
Free PMC Article

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