8TWG

Crystal structure of tetracycline destructase Tet(56-2)

8TWG の概要

• エントリーDOI: 10.2210/pdb8twg/pdb
• 分子名称: 6-hydroxynicotinate 3-monooxygenase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: oxidoreductase
• 由来する生物種: Legionella clemsonensis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 46388.47

構造登録者

Kumar, H.,Tang, W.K.,Tolia, N. (登録日: 2023-08-21, 公開日: 2024-07-03)

主引用文献

Blake, K.S.,Kumar, H.,Loganathan, A.,Williford, E.E.,Diorio-Toth, L.,Xue, Y.P.,Tang, W.K.,Campbell, T.P.,Chong, D.D.,Angtuaco, S.,Wencewicz, T.A.,Tolia, N.H.,Dantas, G.
Sequence-structure-function characterization of the emerging tetracycline destructase family of antibiotic resistance enzymes.
Commun Biol, 7:336-336, 2024

PubMed Abstract: Tetracycline destructases (TDases) are flavin monooxygenases which can confer resistance to all generations of tetracycline antibiotics. The recent increase in the number and diversity of reported TDase sequences enables a deep investigation of the TDase sequence-structure-function landscape. Here, we evaluate the sequence determinants of TDase function through two complementary approaches: (1) constructing profile hidden Markov models to predict new TDases, and (2) using multiple sequence alignments to identify conserved positions important to protein function. Using the HMM-based approach we screened 50 high-scoring candidate sequences in Escherichia coli, leading to the discovery of 13 new TDases. The X-ray crystal structures of two new enzymes from Legionella species were determined, and the ability of anhydrotetracycline to inhibit their tetracycline-inactivating activity was confirmed. Using the MSA-based approach we identified 31 amino acid positions 100% conserved across all known TDase sequences. The roles of these positions were analyzed by alanine-scanning mutagenesis in two TDases, to study the impact on cell and in vitro activity, structure, and stability. These results expand the diversity of TDase sequences and provide valuable insights into the roles of important residues in TDases, and flavin monooxygenases more broadly.

PubMed: 38493211
DOI: 10.1038/s42003-024-06023-w

実験手法

X-RAY DIFFRACTION (1.8 Å)