3FK7

Crystal structure of TetR triple mutant (H64K, S135L, S138I) in complex with 4-ddma-atc

3FK7 の概要

• エントリーDOI: 10.2210/pdb3fk7/pdb
• 関連するPDBエントリー: 2O7O 2TCT 2VKE 2VKV 3FK6
• 分子名称: Tetracycline repressor protein class B from transposon Tn10, Tetracycline repressor protein class D, MAGNESIUM ION, (4aS,12aS)-3,10,11,12a-tetrahydroxy-6-methyl-1,12-dioxo-1,4,4a,5,12,12a-hexahydrotetracene-2-carboxamide, ... (4 entities in total)
• 機能のキーワード: tetracycline repressor, bacterial transcription regulation, altered inducer specificity, 4-de-dimethylamino-anhydrotetracycline, antibiotic resistance, dna-binding, magnesium, metal-binding, repressor, transcription, transcription regulation, transposable element
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 47712.91

構造登録者

Klieber, M.A.,Scholz, O.,Lochner, S.,Gmeiner, P.,Hillen, W.,Muller, Y.A. (登録日: 2008-12-16, 公開日: 2009-11-03, 最終更新日: 2023-11-01)

主引用文献

Klieber, M.A.,Scholz, O.,Lochner, S.,Gmeiner, P.,Hillen, W.,Muller, Y.A.
Structural origins for selectivity and specificity in an engineered bacterial repressor-inducer pair.
Febs J., 276:5610-5621, 2009

PubMed Abstract: The bacterial tetracycline transcription regulation system mediated by the tetracycline repressor (TetR) is widely used to study gene expression in prokaryotes and eukaryotes. To study multiple genes in parallel, a triple mutant TetR(K(64)L(135)I(138)) has been engineered that is selectively induced by the synthetic tetracycline derivative 4-de-dimethylamino-anhydrotetracycline (4-ddma-atc) and no longer by tetracycline, the inducer of wild-type TetR. In the present study, we report the crystal structure of TetR(K(64)L(135)I(138)) in the absence and in complex with 4-ddma-atc at resolutions of 2.1 A. Analysis of the structures in light of the available binding data and previously reported TetR complexes allows for a dissection of the origins of selectivity and specificity. In all crystal structures solved to date, the ligand-binding position, as well as the positioning of the residues lining the binding site, is extremely well conserved, irrespective of the chemical nature of the ligand. Selective recognition of 4-ddma-atc is achieved through fine-tuned hydrogen-bonding constraints introduced by the His64-->Lys substitution, as well as a combination of hydrophobic effect and the removal of unfavorable electrostatic interactions through the introduction of Leu135 and Ile138.

PubMed: 19712110
DOI: 10.1111/j.1742-4658.2009.07254.x

実験手法

X-RAY DIFFRACTION (2.06 Å)