title: The 2.4-A crystal structure of the penicillin-resistant penicillin-binding protein PBP5fm from Enterococcus faecium in complex with benzylpenicillin.
abstract: Penicillin-binding proteins (PBPs) are membrane proteins involved in the final stages of peptidoglycan synthesis and represent the targets of beta-lactam antibiotics. Enterococci are naturally resistant to these antibiotics because they produce a PBP, named PBP5fm in Enterococcus faecium, with low-level affinity for beta-lactams. We report here the crystal structure of the acyl-enzyme complex of PBP5fm with benzylpenicillin at a resolution of 2.4 A. A characteristic of the active site, which distinguishes PBP5fm from other PBPs of known structure, is the topology of the loop 451-465 defining the left edge of the cavity. The residue Arg464, involved in a salt bridge with the residue Asp481, confers a greater rigidity to the PBP5fm active site. In addition, the presence of the Val465 residue, which points into the active site, reducing its accessibility, could account for the low affinity of PBP5fm for beta-lactam. This loop is common to PBPs of low affinity, such as PBP2a from Staphylococcus aureus and PBP3 from Bacillus subtilis. Moreover, the insertion of a serine after residue 466 in the most resistant strains underlines even more the determining role of this loop in the recognition of the substrates.
authors.affiliation: Centre d'Ingénierie des Protéines, Université de Liege, Institut de Physique, Belgium. eric.sauvage@ulg.ac.be
authors.forename: E
authors.initials: E
authors.lastname: Sauvage
authors.forename: F
authors.initials: F
authors.lastname: Kerff
authors.forename: E
authors.initials: E
authors.lastname: Fonzé
authors.forename: R
authors.initials: R
authors.lastname: Herman
authors.forename: B
authors.initials: B
authors.lastname: Schoot
authors.forename: J P
authors.initials: JP
authors.lastname: Marquette
authors.forename: Y
authors.initials: Y
authors.lastname: Taburet
authors.forename: D
authors.initials: D
authors.lastname: Prevost
authors.forename: J
authors.initials: J
authors.lastname: Dumas
authors.forename: G
authors.initials: G
authors.lastname: Leonard
authors.forename: P
authors.initials: P
authors.lastname: Stefanic
authors.forename: J
authors.initials: J
authors.lastname: Coyette
authors.forename: P
authors.initials: P
authors.lastname: Charlier
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018360:Crystallography, X-Ray; D016984:Enterococcus faecium / Q000187:drug effects; D006602:Hexosyltransferases; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D002267:Muramoylpentapeptide Carboxypeptidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D010400:Penicillin G / Q000737:chemistry / Q000378:metabolism*; D010403:Penicillin Resistance; D046915:Penicillin-Binding Proteins; D010406:Penicillins / Q000737:chemistry / Q000378:metabolism*; D010458:Peptidyl Transferases; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D046911:Macromolecular Substances; D046915:Penicillin-Binding Proteins; D010406:Penicillins; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D010400:Penicillin G