Crystal structure of the tet repressor in complex with a novel tetracycline, 9-(N,N-dimethylglycylamido)- 6-demethyl-6-deoxy-tetracycline.

The tetracycline analog 9-(N, N-dimethylglycylamido)-6-demethyl-6-deoxy-tetracycline (9glyTc) belongs to a new group of tetracyclines called glycylcyclines. They are strong antibiotics showing reduced sensitivity against the major tetracycline resistance mechanisms. We have determined the crystal structure of 9glyTc in complex with Tet repressor class D, TetR(D), at 2.4 A resolution. Sterical hindrance at the entrance of the tetracycline binding tunnel of TetR by the bulky and charged glycyl amido substituent interferes with conformational changes required for the mechanism of induction, and leads to decreased induction efficiency as observed for point mutations of amino acid residues located in the neighbourhood to the glycylamido moiety of bound 9glyTc.

[1]  F. Cordes,et al.  Conformational changes of the Tet repressor induced by tetracycline trapping. , 1998, Journal of molecular biology.

[2]  W. Hillen,et al.  Fast large-scale purification of tetracycline repressor variants from overproducing Escherichia coli strains. , 1996, Journal of chromatography. A.

[3]  W. Hillen,et al.  Tetracycline analogs affecting binding to Tn10-Encoded Tet repressor trigger the same mechanism of induction. , 1996, Biochemistry.

[4]  D. A. Dougherty,et al.  Cation-π Interactions in Chemistry and Biology: A New View of Benzene, Phe, Tyr, and Trp , 1996, Science.

[5]  W. Saenger,et al.  Characterization of non-inducible Tet repressor mutants suggests conformational changes necessary for induction , 1995, Nature Structural Biology.

[6]  W. Saenger,et al.  The complex formed between Tet repressor and tetracycline-Mg2+ reveals mechanism of antibiotic resistance. , 1995 .

[7]  A. Yamaguchi,et al.  A novel glycylcycline, 9-(N,N-dimethylglycylamido)-6-demethyl-6-deoxytetracycline, is neither transported nor recognized by the transposon Tn10-encoded metal-tetracycline/H+ antiporter , 1995, Antimicrobial agents and chemotherapy.

[8]  E A Merritt,et al.  Raster3D Version 2.0. A program for photorealistic molecular graphics. , 1994, Acta crystallographica. Section D, Biological crystallography.

[9]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[10]  W. Saenger,et al.  Structure of the Tet repressor-tetracycline complex and regulation of antibiotic resistance. , 1994, Science.

[11]  J. Bloom,et al.  Glycylcyclines. 1. A new generation of potent antibacterial agents through modification of 9-aminotetracyclines. , 1994, Journal of medicinal chemistry.

[12]  N. Jacobus,et al.  In vitro and in vivo antibacterial activities of the glycylcyclines, a new class of semisynthetic tetracyclines , 1993, Antimicrobial Agents and Chemotherapy.

[13]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[14]  A. Salyers,et al.  Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance , 1992, Clinical Microbiology Reviews.

[15]  H. Neu,et al.  The Crisis in Antibiotic Resistance , 1992, Science.

[16]  W. Hillen,et al.  The role of the N terminus in Tet repressor for tet operator binding determined by a mutational analysis. , 1992, The Journal of biological chemistry.

[17]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[18]  W. Hillen,et al.  Structural requirements of tetracycline-Tet repressor interaction: determination of equilibrium binding constants for tetracycline analogs with the Tet repressor , 1991, Antimicrobial Agents and Chemotherapy.

[19]  W. Hillen,et al.  Selection for Tn10 tet repressor binding to tet operator in Escherichia coli: isolation of temperature-sensitive mutants and combinatorial mutagenesis in the DNA binding motif. , 1991, Genetics.

[20]  V. Luzzati,et al.  Traitement statistique des erreurs dans la determination des structures cristallines , 1952 .

[21]  W. Hillen,et al.  Mechanisms underlying expression of Tn10 encoded tetracycline resistance. , 1994, Annual review of microbiology.

[22]  Axel T. Brunger,et al.  Simulated Annealing in Crystallography , 1991 .