Enolpyruvate transferase MurAAA149E, identified during adaptation of Enterococcus faecium to daptomycin, increases stability of MurAA–MurG interaction

Yue Zhou, Budi Utama, Shivendra Pratap, Adeline Supandy, Xinhao Song, Truc T. Tran, Heer H. Mehta, Cesar A. Arias, Yousif Shamoo

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


Daptomycin (DAP) is an antibiotic frequently used as a drug of last resort against vancomycin-resistant enterococci. One of the major challenges when using DAP against vancomycin-resistant enterococci is the emergence of resistance, which is mediated by the cell-envelope stress system LiaFSR. Indeed, inhibition of LiaFSR signaling has been suggested as a strategy to “resensitize” enterococci to DAP. In the absence of LiaFSR, alternative pathways mediating DAP resistance have been identified, including adaptive mutations in the enolpyruvate transferase MurAA (MurAAA149E), which catalyzes the first committed step in peptidoglycan biosynthesis; however, how these mutations confer resistance is unclear. Here, we investigated the biochemical basis for MurAAA149E-mediated adaptation to DAP to determine whether such an alternative pathway would undermine the potential efficacy of therapies that target the LiaFSR pathway. We found cells expressing MurAAA149E had increased susceptibility to glycoside hydrolases, consistent with decreased cell wall integrity. Furthermore, structure–function studies of MurAA and MurAAA149E using X-ray crystallography and biochemical analyses indicated only a modest decrease in MurAAA149E activity, but a 16-fold increase in affinity for MurG, which performs the last intracellular step of peptidoglycan synthesis. Exposure to DAP leads to mislocalization of cell division proteins including MurG. In Bacillus subtilis, MurAA and MurG colocalize at division septa and, thus, we propose MurAAA149E may contribute to DAP nonsusceptibility by increasing the stability of MurAA–MurG interactions to reduce DAP-induced mislocalization of these essential protein complexes.

Original languageEnglish (US)
Article number102912
JournalJournal of Biological Chemistry
Issue number3
StatePublished - Mar 2023


  • Enterococcus
  • UDP-N-acetylglucosamine enolpyruvyl transferase
  • X-ray crystallography
  • antibiotic resistance
  • enzyme kinetics
  • immunofluorescence microscopy
  • peptidoglycan
  • Transferases/metabolism
  • Drug Resistance, Bacterial
  • Anti-Bacterial Agents/pharmacology
  • Enterococcus faecium/drug effects
  • Microbial Sensitivity Tests
  • Bacterial Proteins/metabolism
  • Daptomycin/metabolism
  • Peptidoglycan/metabolism

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry
  • Cell Biology


Dive into the research topics of 'Enolpyruvate transferase MurAAA149E, identified during adaptation of Enterococcus faecium to daptomycin, increases stability of MurAA–MurG interaction'. Together they form a unique fingerprint.

Cite this