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Antimicrobial Agents and Chemotherapy, April 2001, p. 1053-1057, Vol. 45, No. 4
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.4.1053-1057.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Peptide Deformylase as an Antibacterial Drug Target: Assays for Detection of Its Inhibition in Escherichia coli Cell Homogenates and Intact Cells

Christian M. Apfel,* Stefan Evers, Christian Hubschwerlen,dagger Wolfgang Pirson, Malcolm G. P. Page, and Wolfgang Keckdagger

Pharma Research Basel, F. Hoffmann-La Roche Ltd., CH-4070 Basel, Switzerland

Received 10 August 2000/Returned for modification 5 October 2000/Accepted 20 December 2000

An assay was developed to determine the activity of peptide deformylase (PDF) inhibitors under conditions as close as possible to the physiological situation. The assay principle is the detection of N-terminal [35S]methionine labeling of a protein that contains no internal methionine. If PDF is active, the deformylation of the methionine renders the peptide a substrate for methionine aminopeptidase, resulting in the removal of the N-terminal methionine label. In the presence of a PDF inhibitor, the deformylation is blocked so that the N-formylated peptide is not processed and the label is detected. Using this assay, it is possible to determine the PDF activity under near-physiological conditions in a cell-free transcription-translation system as well as in intact bacterial cells.

* Corresponding author. Mailing address: F. Hoffmann-La Roche Ltd., PRBM-H, Bldg. 69/11A, CH-4070 Basel, Switzerland. Phone: 41 61 688 5878. Fax: 41 61 688 2377. E-mail: christian.apfel{at}roche.com.

dagger Present address: Morphochem AG, Basel, Switzerland.

Antimicrobial Agents and Chemotherapy, April 2001, p. 1053-1057, Vol. 45, No. 4
0066-4804/01/$04.00+0   DOI: 10.1128/AAC.45.4.1053-1057.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Watters, A. A., Jones, R. N., Leeds, J. A., Denys, G., Sader, H. S., Fritsche, T. R. (2006). Antimicrobial activity of a novel peptide deformylase inhibitor, LBM415, tested against respiratory tract and cutaneous infection pathogens: a global surveillance report (2003-2004). J Antimicrob Chemother 57: 914-923 [Abstract] [Full Text]  
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  • Apfel, C. M., Locher, H., Evers, S., Takács, B., Hubschwerlen, C., Pirson, W., Page, M. G. P., Keck, W. (2001). Peptide Deformylase as an Antibacterial Drug Target: Target Validation and Resistance Development. Antimicrob. Agents Chemother. 45: 1058-1064 [Abstract] [Full Text]  


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