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Antimicrobial Agents and Chemotherapy, August 2005, p. 3367-3372, Vol. 49, No. 8
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.8.3367-3372.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Fluorescence Polarization Method To Characterize Macrolide-Ribosome Interactions

Kang Yan,1* Eric Hunt,2 John Berge,2 Earl May,1 Robert A. Copeland,1 and Richard R. Gontarek1

Department of Enzymology and Mechanistic Pharmacology, GlaxoSmithKline, 1250 South Collegeville Road, Collegeville, Pennsylvania 19426,1 Department of Medicinal Chemistry, GlaxoSmithKline, New Frontiers Science Park, Harlow, CM19 5AW, United Kingdom2

Received 12 March 2005/ Returned for modification 2 April 2005/ Accepted 7 May 2005

A fluorescence polarization assay is described that measures the binding of fluorescently labeled erythromycin to 70S ribosomes from Escherichia coli and the displacement of the erythromycin from these ribosomes. The assay has been validated with several macrolide derivatives and other known antibiotics. We demonstrate that this assay is suitable for determining the dissociation constants of novel compounds that have binding sites overlapping those of macrolides. This homogeneous binding assay provides a valuable tool for defining structure-activity relationships among compounds during the discovery and development of new ribosome-targeting drugs.

* Corresponding author. Mailing address: 1250 South Collegeville Road, Collegeville, PA 19426. Phone: (610) 917-7282. Fax: (610) 917-7901. E-mail: kang.2.yan{at}gsk.com.

Antimicrobial Agents and Chemotherapy, August 2005, p. 3367-3372, Vol. 49, No. 8
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.8.3367-3372.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Davidovich, C., Bashan, A., Auerbach-Nevo, T., Yaggie, R. D., Gontarek, R. R., Yonath, A. (2007). Induced-fit tightens pleuromutilins binding to ribosomes and remote interactions enable their selectivity. Proc. Natl. Acad. Sci. USA 104: 4291-4296 [Abstract] [Full Text]  
  • Yan, K., Madden, L., Choudhry, A. E., Voigt, C. S., Copeland, R. A., Gontarek, R. R. (2006). Biochemical Characterization of the Interactions of the Novel Pleuromutilin Derivative Retapamulin with Bacterial Ribosomes. Antimicrob. Agents Chemother. 50: 3875-3881 [Abstract] [Full Text]  


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