Structure and function of the glycopeptide N-methyltransferase MtfA, a tool for the biosynthesis of modified glycopeptide antibiotics

Chem Biol. 2009 Apr 24;16(4):401-10. doi: 10.1016/j.chembiol.2009.02.007.

Abstract

There is a considerable interest in the modification of existing antibiotics to generate new antimicrobials. Glycopeptide antibiotics (GPAs) are effective against serious Gram-positive bacterial pathogens including methicillin-resistant Staphylococcus aureus. However, resistance to these antibiotics is becoming a serious problem requiring new strategies. We show that the Amycolatopsis orientalis (S)-adenosyl-L-methionine-dependent methyltransferase MtfA, from the vancomycin-class GPA chloroeremomycin biosynthetic pathway, catalyzes in vivo and in vitro methyl transfer to generate methylated GPA derivatives of the teicoplanin class. The crystal structure of MtfA complexed with (S)-adenosyl-L-methionine, (S)-adenosylhomocysteine, or sinefungin inhibitor, coupled with mutagenesis, identified His228 as a likely general base required for methyl transfer to the N terminus of the glycopeptide. Computational docking and molecular dynamics simulations were used to model binding of demethyl-vancomycin aglycone to MtfA. These results demonstrate its utility as a tool for engineering methylated analogs of GPAs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actinomycetales / enzymology*
  • Adenosine / analogs & derivatives
  • Adenosine / chemistry
  • Adenosine / metabolism
  • Anti-Bacterial Agents / chemistry
  • Anti-Bacterial Agents / metabolism*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Catalytic Domain
  • Crystallography, X-Ray
  • Methyltransferases / chemistry*
  • Methyltransferases / genetics
  • Methyltransferases / metabolism*
  • Models, Molecular
  • Point Mutation
  • Protein Binding
  • Protein Multimerization
  • S-Adenosylhomocysteine / chemistry
  • S-Adenosylhomocysteine / metabolism
  • S-Adenosylmethionine / chemistry
  • S-Adenosylmethionine / metabolism
  • Teicoplanin / metabolism

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Teicoplanin
  • S-Adenosylmethionine
  • S-Adenosylhomocysteine
  • Methyltransferases
  • S-adenosyl-L-methionine-dependent N-methyltransferase
  • Adenosine
  • sinefungin

Associated data

  • PDB/3G2M
  • PDB/3G2O
  • PDB/3G2P
  • PDB/3G2Q