This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services E-mail this article to a friend Similar articles in this journal Similar articles in ASM journals Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Degenkolb, J Articles by Hillen, W Search for Related Content PubMed PubMed Citation Articles by Degenkolb, J Articles by Hillen, W

 Previous Article  |  Next Article 

Antimicrob Agents Chemother. 1991 August; 35(8): 1591-1595

Structural requirements of tetracycline-Tet repressor interaction: determination of equilibrium binding constants for tetracycline analogs with the Tet repressor.

J Degenkolb, M Takahashi, G A Ellestad and W Hillen

Lehrstuhl für Mikrobiologie, Friedrich-Alexander Universität Erlangen-Nürnberg, Federal Republic of Germany.

ABSTRACT

We used the Tn10-encoded Tet repressor, which has a highly specific binding capacity for tetracycline, to probe contacts between the drug and protein by chemical interference studies of the antibiotic. For that purpose, the equilibrium association constants of modified tetracyclines with the Tet repressor and Mg2+ cations were determined quantitatively. The results confirm the previous notion that Mg2+ probably binds with the oxygens at positions 11 and 12 and is absolutely required for protein-drug recognition. Modifications were introduced at positions seven, six, five, and four of the drug, and anhydrotetracycline was also studied. Substitutions or eliminations of functions at these positions influenced binding to the Tet repressor up to 35-fold. The introduction of an azido function at position seven in 7-azidotetracycline and epimerization of the substituents at position four in 4-epitetracycline lead to a 2- or 25-fold reduction, respectively, of Tet repressor affinity in those compounds. Anhydrotetracycline bound about 35-fold more strongly than tetracycline did, indicating that the oxygen at position 11 may be involved in Tet repressor recognition. This increased binding is in contrast to the lower antibiotic activity of anhydrotetracycline and indicates that the Tet repressor and ribosomes recognize the drug differently.

---

Antimicrob Agents Chemother. 1991 August; 35(8): 1591-1595

This article has been cited by other articles:

• Stary, E., Gaupp, R., Lechner, S., Leibig, M., Tichy, E., Kolb, M., Bertram, R. (2010). New Architectures for Tet-On and Tet-Off Regulation in Staphylococcus aureus. Appl. Environ. Microbiol. 76: 680-687 [Abstract] [Full Text]  
• Klotzsche, M., Berens, C., Hillen, W. (2005). A Peptide Triggers Allostery in Tet Repressor by Binding to a Unique Site. J. Biol. Chem. 280: 24591-24599 [Abstract] [Full Text]  
• Biburger, M., Berens, C., Lederer, T., Krec, T., Hillen, W. (1998). Intragenic Suppressors of Induction-Deficient TetR Mutants: Localization and Potential Mechanism of Action. J. Bacteriol. 180: 737-741 [Abstract] [Full Text]  
• Berens, C., Schnappinger, D., Hillen, W. (1997). The Role of the Variable Region in Tet Repressor for Inducibility by Tetracycline. J. Biol. Chem. 272: 6936-6942 [Abstract] [Full Text]  
• Hinrichs, W, Kisker, C, Duvel, M, Muller, A, Tovar, K, Hillen, W, Saenger, W (1994). Structure of the Tet repressor-tetracycline complex and regulation of antibiotic resistance. Science 264: 418-420 [Abstract]  
• Urlinger, S., Baron, U., Thellmann, M., Hasan, M. T., Bujard, H., Hillen, W. (2000). Exploring the sequence space for tetracycline-dependent transcriptional activators: Novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. USA 97: 7963-7968 [Abstract] [Full Text]