5UXC

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

Summary for 5UXC

• Entry DOI: 10.2210/pdb5uxc/pdb
• Related: 5UXA 5UXB 5UXD
• Descriptor: Predicted aminoglycoside phosphotransferase, MAGNESIUM ION, GUANOSINE-5'-DIPHOSPHATE, ... (10 entities in total)
• Functional Keywords: 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
• Biological source: Brachybacterium faecium
• Total number of polymer chains: 1
• Total formula weight: 34951.27

Authors

Stogios, P.J.,Skarina, T.,Wawrzak, Z.,Yim, V.,Savchenko, A.,Anderson, W.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (deposition date: 2017-02-22, release date: 2017-08-16, Last modification date: 2024-10-16)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.72 Å)