title: A PBP2x from a clinical isolate of Streptococcus pneumoniae exhibits an alternative mechanism for reduction of susceptibility to beta-lactam antibiotics.
abstract: The human pathogen Streptococcus pneumoniae is one of the main causative agents of respiratory tract infections. At present, clinical isolates of S. pneumoniae often exhibit decreased susceptibility toward beta-lactams, a phenomenon linked to multiple mutations within the penicillin-binding proteins (PBPs). PBP2x, one of the six PBPs of S. pneumoniae, is the first target to be modified under antibiotic pressure. By comparing 89 S. pneumoniae PBP2x sequences from clinical and public data bases, we have identified one major group of sequences from drug-sensitive strains as well as two distinct groups from drug-resistant strains. The first group includes proteins that display high similarity to PBP2x from the well characterized resistant strain Sp328. The second group includes sequences in which a signature mutation, Q552E, is found adjacent to the third catalytic motif. In this work, a PBP2x from a representative strain from the latter group (S. pneumoniae 5259) was biochemically and structurally characterized. Phenotypical analyses of transformed pneumococci show that the Q552E substitution is responsible for most of the reduction of strain susceptibility toward beta-lactams. The crystal structure of 5259-PBP2x reveals a change in polarity and charge distribution around the active site cavity, as well as rearrangement of strand beta3, emulating structural changes observed for other PBPs that confer drug resistance to Gram-positive pathogens. Interestingly, the active site of 5259-PBP2x is in closed conformation, whereas that of Sp328-PBP2x is open. Consequently, S. pneumoniae has evolved to employ the same protein in two distinct mechanisms of antibiotic resistance.
authors.affiliation: Laboratoire de Cristallographie Macromoléculaire, Institut de Biologie Structurale Jean-Pierre Ebel (CNRS/CEA/UJF), Grenoble, France.
authors.forename: Lucile
authors.initials: L
authors.lastname: Pernot
authors.forename: Laurent
authors.initials: L
authors.lastname: Chesnel
authors.forename: Audrey
authors.initials: A
authors.lastname: Le Gouellec
authors.forename: Jacques
authors.initials: J
authors.lastname: Croizé
authors.forename: Thierry
authors.initials: T
authors.lastname: Vernet
authors.forename: Otto
authors.initials: O
authors.lastname: Dideberg
authors.forename: Andréa
authors.initials: A
authors.lastname: Dessen
mesh_terms: D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D046915:Penicillin-Binding Proteins; D011487:Protein Conformation; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018440:beta-Lactam Resistance / Q000235:genetics*
chemical_list: D002352:Carrier Proteins; D046915:Penicillin-Binding Proteins; D012261:Ribonucleoproteins; C072223:PBP 2x protein, Streptococcus