title: Role of penicillin-binding protein 5 C-terminal amino acid substitutions in conferring ampicillin resistance in Norwegian clinical strains of Enterococcus faecium.
abstract: The importance of amino acid sequence differences in the C-terminal part and levels of mRNA expression of penicillin-binding protein 5 (PBP5) for ampicillin resistance in Enterococcus faecium was investigated. Seventeen isolates from Norwegian hospitalized patients (ampicillin MIC 0.064->256 mg/L) with different C-terminal pbp5 DNA sequences encoding 11 different amino acid sequences were analyzed with a 14C-radiolabeled penicillin- binding assay to PBP5 and with real-time PCR quantification of pbp5 mRNA expression. Using multiple logistic regression analysis the amino acid substitution Met 485 was linked to ampicillin MIC and levels of 14C-radiolabeled penicillin bound to PBP5; however, there were isolates with identical PBP5 alleles and different ampicillin MICs. There was no relation between the quantity of pbp5 mRNA transcripts and ampicillin resistance. The results cannot explain ampicillin resistance in Norwegian clinical strains of E. faecium and indicate that other factors besides the properties of the C-terminal part of PBP5 are most likely involved.
authors.affiliation: Institute of Medicine, University of Bergen, Haukeland University Hospital, Norway. roland.jureen@med.uib.no
authors.forename: R
authors.initials: R
authors.lastname: Jureen
authors.forename: S C
authors.initials: SC
authors.lastname: Mohn
authors.forename: S
authors.initials: S
authors.lastname: Harthug
authors.forename: L
authors.initials: L
authors.lastname: Haarr
authors.forename: N
authors.initials: N
authors.lastname: Langeland
mesh_terms: D019943:Amino Acid Substitution; D000668:Ampicillin Resistance / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D017931:DNA Primers; D016984:Enterococcus faecium / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D006602:Hexosyltransferases / Q000235:genetics*; D006801:Humans; D015999:Multivariate Analysis; D002267:Muramoylpentapeptide Carboxypeptidase / Q000235:genetics*; D009664:Norway; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D012007:Rectum / Q000382:microbiology; D001618:beta-Lactamases / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D046915:Penicillin-Binding Proteins; D012329:RNA, Bacterial; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D001618:beta-Lactamases