Tetracycline does not directly inhibit the function of bacterial elongation factor Tu
Gzyl, Katherine E.
Public Library of Science
Understanding the molecular mechanism of antibiotics that are currently in use is important for the development of new antimicrobials. The tetracyclines, discovered in the 1940s, are a well-established class of antibiotics that still have a role in treating microbial infections in humans. It is generally accepted that the main target of their action is the ribosome. The esti- mated affinity for tetracycline binding to the ribosome is relatively low compared to the actual potency of the drug in vivo . Therefore, additional inhibitory effects of tetracycline on the translation machinery have been discussed. Structural evidence suggests that tetracycline inhibits the function of the essential bacterial GTPase Elongation Factor (EF)-Tu through interaction with the bound nucleotide. Based on this, tetracycline has been predicted to impede the nucleotide-binding properties of EF-Tu. However, detailed kinetic studies addressing the effect of tetracycline on nucleotide binding have been prevented by the fluorescence properties of the antibiotic. Here, we report a fluorescence-bas ed kinetic assay that minimizes the effect of tetracycline autofluorescence, enabling the detailed kinetic analysis of the nucleotide-bin ding properties of Escherichia coli EF-Tu. Further- more, using physiologica lly relevant conditions, we demonstrate that tetracycline does not affect EF-Tu’s intrinsic or ribosome-stimulated GTPase activity, nor the stability of the EF- Tu•GTP•Phe-tRNA Phe complex. We therefore provide clear evidence that tetracycline does not directly impede the function of EF-Tu.
Sherpa Romeo green journal. Open access article. Creative Commons Attribution 4.0 International License (CC BY 4.0) applies.
Tetracycline , EF-Tu , Elongation factor , Kinetic analysis , Nucleotide binding , Autofluorescence , GTPase , Inhibit , Antibiotic , Antimicrobial drugs
Gzyl, K.E., & Wieden, H-J. (2017). Tetracycline does not directly inhibit the function of bacterial elongation factor Tu. PloS ONE, 12(5), e0178523. https://doi.org/10.1371/journal.pone.0178523