scholarly journals Comparative analysis of Pseudomonas aeruginosa penicillin-binding protein 7 in the context of its membership in the family of low-molecular-mass PBPs

Microbiology ◽  
1998 ◽  
Vol 144 (4) ◽  
pp. 975-983 ◽  
Author(s):  
J. Song ◽  
G. Xie ◽  
P. K. Elf ◽  
K. D. Young ◽  
R. A. Jensen
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Comparative Analysis ◽  
Molecular Mass ◽  
Binding Protein ◽  
Penicillin Binding Protein ◽  
The Family

scholarly journals Structural Insights into Ceftobiprole Inhibition of Pseudomonas aeruginosa Penicillin-Binding Protein 3

2020 ◽  
Vol 64 (5) ◽  
Author(s):  
Vijay Kumar ◽  
Christie Tang ◽  
Christopher R. Bethel ◽  
Krisztina M. Papp-Wallace ◽  
Jacob Wyatt ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Binding Sites ◽  
Binding Protein ◽  
Covalent Bond ◽  
Penicillin Binding Protein ◽  
Content Type ◽  
Cephalosporin Antibiotic ◽  
Structural Insights ◽  
Fine Tune ◽  
Advanced Generation

ABSTRACT Ceftobiprole is an advanced-generation broad-spectrum cephalosporin antibiotic with potent and rapid bactericidal activity against Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus, as well as susceptible Gram-negative pathogens, including Pseudomonas sp. pathogens. In the case of Pseudomonas aeruginosa, ceftobiprole acts by inhibiting P. aeruginosa penicillin-binding protein 3 (PBP3). Structural studies were pursued to elucidate the molecular details of this PBP inhibition. The crystal structure of the His-tagged PBP3-ceftobiprole complex revealed a covalent bond between the ligand and the catalytic residue S294. Ceftobiprole binding leads to large active site changes near binding sites for the pyrrolidinone and pyrrolidine rings. The S528 to L536 region adopts a conformation previously not observed in PBP3, including partial unwinding of the α11 helix. These molecular insights can lead to a deeper understanding of β-lactam-PBP interactions that result in major changes in protein structure, as well as suggesting how to fine-tune current inhibitors and to develop novel inhibitors of this PBP.

scholarly journals Pan-β-Lactam Resistance Development in Pseudomonas aeruginosa Clinical Strains: Molecular Mechanisms, Penicillin-Binding Protein Profiles, and Binding Affinities

2012 ◽  
Vol 56 (9) ◽  
pp. 4771-4778 ◽  
Author(s):  
Bartolomé Moyá ◽  
Alejandro Beceiro ◽  
Gabriel Cabot ◽  
Carlos Juan ◽  
Laura Zamorano ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Molecular Mechanisms ◽  
Efflux Pump ◽  
Binding Protein ◽  
Efflux Pumps ◽  
Binding Affinities ◽  
Penicillin Binding Protein ◽  
Content Type ◽  
Reverse Transcription Pcr

ABSTRACTWe investigated the mechanisms leading toPseudomonas aeruginosapan-β-lactam resistance (PBLR) development during the treatment of nosocomial infections, with a particular focus on the modification of penicillin-binding protein (PBP) profiles and imipenem, ceftazidime, and ceftolozane (former CXA-101) PBP binding affinities. For this purpose, six clonally related pairs of sequential susceptible-PBLR isolates were studied. The presence ofoprD,ampD, anddacBmutations was explored by PCR followed by sequencing and the expression ofampCand efflux pump genes by real-time reverse transcription-PCR. The fluorescent penicillin Bocillin FL was used to determine PBP profiles in membrane preparations from all pairs, and 50% inhibitory concentrations (IC50s) of ceftolozane, ceftazidime, and imipenem were analyzed in 3 of them. Although a certain increase was noted (0 to 5 2-fold dilutions), the MICs of ceftolozane were ≤4 μg/ml in all PBLR isolates. All 6 PBLR isolates lacked OprD and overexpressedampCand one or several efflux pumps, particularlymexBand/ormexY. Additionally, 5 of them showed modified PBP profiles, including a modified pattern (n= 1) or diminished expression (n= 1) of PBP1a and a lack of PBP4 expression (n= 4), which correlated with AmpC overexpression driven bydacBmutation. Analysis of the essential PBP IC50s revealed significant variation of PBP1a/b binding affinities, both within each susceptible-PBLR pair and across the different pairs. Moreover, despite the absence of significant differences in gene expression or sequence, a clear tendency toward increased PBP2 (imipenem) and PBP3 (ceftazidime, ceftolozane, imipenem) IC50s was noted in PBLR isolates. Thus, our results suggest that in addition to AmpC, efflux pumps, and OprD, the modification of PBP patterns appears to play a role in thein vivoemergence of PBLR strains, which still conserve certain susceptibility to the new antipseudomonal cephalosporin ceftolozane.

scholarly journals Affinity of Tomopenem (CS-023) for Penicillin-Binding Proteins in Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa

2008 ◽  
Vol 53 (3) ◽  
pp. 1238-1241 ◽  
Author(s):  
Tetsufumi Koga ◽  
Chika Sugihara ◽  
Masayo Kakuta ◽  
Nobuhisa Masuda ◽  
Eiko Namba ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Binding Proteins ◽  
Binding Protein ◽  
Penicillin Binding Protein ◽  
Penicillin Binding Proteins ◽  
E Coli ◽  
High Affinity

ABSTRACT Tomopenem (formerly CS-023), a novel 1β-methylcarbapenem, exhibited high affinity for penicillin-binding protein (PBP) 2 in Staphylococcus aureus, PBP 2 in Escherichia coli, and PBPs 2 and 3 in Pseudomonas aeruginosa, which are considered major lethal targets. Morphologically, tomopenem induced spherical forms in E. coli and short filamentation with bulges in P. aeruginosa, which correlated with the drug's PBP profiles. The potential of resistance of these bacteria to tomopenem was comparable to that to imipenem.

Production and purification of the penicillin-binding protein 3 from Pseudomonas aeruginosa

2010 ◽  
Vol 73 (2) ◽  
pp. 177-183 ◽  
Author(s):  
Sandra Ruiloba de León ◽  
Kathy Daniels ◽  
Anthony J. Clarke
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Binding Protein ◽  
Penicillin Binding Protein

scholarly journals Cloning and Characterization of a Gene,pbpF, Encoding a New Penicillin-Binding Protein, PBP2B, inStaphylococcus aureus

1999 ◽  
Vol 43 (7) ◽  
pp. 1578-1583 ◽  
Author(s):  
Hitoshi Komatsuzawa ◽  
Gil H. Choi ◽  
Kouji Ohta ◽  
Motoyuki Sugai ◽  
Monique T. Tran ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Binding Protein ◽  
Protein Band ◽  
Binding Activity ◽  
Amino Acid Residues ◽  
Penicillin Binding Protein ◽  
Putative Signal Peptide ◽  
C Terminus ◽  
Histidine Tag

ABSTRACT A previously unrecognized penicillin binding protein (PBP) gene,pbpF, was identified in Staphylococcus aureus. This gene encodes a protein of 691 amino acid residues with an estimated molecular mass of 78 kDa. The molecular mass is very close to that of S. aureus PBP2 (81 kDa), and the protein is tentatively named PBP2B. PBP2B has three motifs, SSVK, SSN, and KTG, that can be found in PBPs and β-lactamases. Recombinant PBP2B (rPBP2B), which lacks a putative signal peptide at the N terminus and has a histidine tag at the C terminus, was expressed inEscherichia coli. The purified rPBP2B was shown to have penicillin binding activity. A protein band was detected from S. aureus membrane fraction by immunoblotting with anti-rPBP2B serum. Also, penicillin binding activity of the protein immunoprecipitated with anti-rPBP2B serum was detected. These results suggest the presence of PBP2B in S. aureus cell membrane that covalently binds penicillin. The internal region ofpbpF and PBP2B protein were found in all 12 S. aureus strains tested by PCR and immunoblotting.

Sign in / Sign up

Export Citation Format

Share Document