5UXD

Crystal structure of macrolide 2'-phosphotransferase MphH from Brachybacterium faecium in complex with azithromycin

5UXD の概要

• エントリーDOI: 10.2210/pdb5uxd/pdb
• 関連するPDBエントリー: 5UXA 5UXB 5UXC
• 分子名称: Macrolide 2'-phosphotransferase MphH, AZITHROMYCIN, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: antibiotic resistance, macrolide, cave bacterium, phosphotransferase, kinase, alpha/beta protein, azithromycin, structural genomics, center for structural genomics of infectious diseases, csgid, national institute of allergy and infectious diseases, niaid, transferase-antibiotic complex, transferase/antibiotic
• 由来する生物種: Brachybacterium faecium (strain ATCC 43885 / DSM 4810 / NCIB 9860)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 67078.09

構造登録者

Stogios, P.J.,Skarina, T.,Wawrzak, Z.,Yim, V.,Savchenko, A.,Anderson, W.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (登録日: 2017-02-22, 公開日: 2017-08-23, 最終更新日: 2023-10-04)

主引用文献

Pawlowski, A.C.,Stogios, P.J.,Koteva, K.,Skarina, T.,Evdokimova, E.,Savchenko, A.,Wright, G.D.
The evolution of substrate discrimination in macrolide antibiotic resistance enzymes.
Nat Commun, 9:112-112, 2018

PubMed Abstract: The production of antibiotics by microbes in the environment and their use in medicine and agriculture select for existing and emerging resistance. To address this inevitability, prudent development of antibiotic drugs requires careful consideration of resistance evolution. Here, we identify the molecular basis for expanded substrate specificity in MphI, a macrolide kinase (Mph) that does not confer resistance to erythromycin, in contrast to other known Mphs. Using a combination of phylogenetics, drug-resistance phenotypes, and in vitro enzyme assays, we find that MphI and MphK phosphorylate erythromycin poorly resulting in an antibiotic-sensitive phenotype. Using likelihood reconstruction of ancestral sequences and site-saturation combinatorial mutagenesis, supported by Mph crystal structures, we determine that two non-obvious mutations in combination expand the substrate range. This approach should be applicable for studying the functional evolution of any antibiotic resistance enzyme and for evaluating the evolvability of resistance enzymes to new generations of antibiotic scaffolds.

PubMed: 29317655
DOI: 10.1038/s41467-017-02680-0

実験手法

X-RAY DIFFRACTION (1.7 Å)