title: Crystal structures of penicillin-binding protein 3 in complexes with azlocillin and cefoperazone in both acylated and deacylated forms.
abstract: Penicillin-binding protein 3 (PBP3) from Pseudomonas aeruginosa is the molecular target of β-lactam-based antibiotics. Structures of PBP3 in complexes with azlocillin and cefoperazone, which are in clinical use for the treatment of pseudomonad infections, have been determined to 2.0 Å resolution. Together with data from other complexes, these structures identify a common set of residues involved in the binding of β-lactams to PBP3. Comparison of wild-type and an active site mutant (S294A) showed that increased thermal stability of PBP3 following azlocillin binding was entirely due to covalent binding to S294, whereas cefoperazone binding produces some increase in stability without the covalent link. Consistent with this, a third crystal structure was determined in which the hydrolysis product of cefoperazone was noncovalently bound in the active site of PBP3. This is the first structure of a complex between a penicillin-binding protein and cephalosporic acid and may be important in the design of new noncovalent PBP3 inhibitors.
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK.
authors.forename: Jingshan
authors.initials: J
authors.lastname: Ren
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK.|OPPF-UK, The Research Complex at Harwell, Rutherford Appleton Laboratory, Oxfordshire, UK.
authors.forename: Joanne E
authors.initials: JE
authors.lastname: Nettleship
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK.|OPPF-UK, The Research Complex at Harwell, Rutherford Appleton Laboratory, Oxfordshire, UK.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Males
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK.|Diamond Light Sources, Harwell Science and Innovation Campus, Didcot, UK.
authors.forename: David I
authors.initials: DI
authors.lastname: Stuart
authors.affiliation: Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Oxford, UK.|OPPF-UK, The Research Complex at Harwell, Rutherford Appleton Laboratory, Oxfordshire, UK.
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Owens
mesh_terms: D000215:Acylation; D001390:Azlocillin / Q000737:chemistry*; D002438:Cefoperazone / Q000737:chemistry*; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D015394:Molecular Structure; D046915:Penicillin-Binding Proteins / Q000737:chemistry*
chemical_list: D046915:Penicillin-Binding Proteins; D002438:Cefoperazone; D001390:Azlocillin
keywords: Pseudomonas aeruginosa; azlocillin; cefoperazone; penicillin-binding protein; thermal shift assay; β-lactam antibiotics
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 21;107(51):22002-7
references.pmid: 21135211
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):235-42
references.pmid: 21460441
references.citation: Antimicrob Agents Chemother. 2012 Sep;56(9):4771-8
references.pmid: 22733064
references.citation: J Mol Biol. 2012 Oct 26;423(3):351-64
references.pmid: 22846910
references.citation: Molecules. 2012;17(11):12478-505
references.pmid: 23095893
references.citation: Ann N Y Acad Sci. 2013 Jan;1277:76-83
references.pmid: 23346858
references.citation: ACS Chem Biol. 2013 Oct 18;8(10):2112-6
references.pmid: 23899657
references.citation: PLoS One. 2014;9(5):e98042
references.pmid: 24875494
references.citation: Antimicrob Agents Chemother. 1979 Aug;16(2):150-7
references.pmid: 314775
references.citation: J Mol Biol. 2011 Jan 7;405(1):173-84
references.pmid: 20974151
references.citation: Antimicrob Agents Chemother. 2010 Sep;54(9):3933-7
references.pmid: 20547785
references.citation: Protein Expr Purif. 2010 Oct;73(2):177-83
references.pmid: 20580675
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):426-36
references.pmid: 20382996
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
references.pmid: 20124702
references.citation: Methods Mol Biol. 2008;426:299-318
references.pmid: 18542872
references.citation: Respir Care. 2008 Apr;53(4):471-9
references.pmid: 18364060
references.citation: FEMS Microbiol Rev. 2008 Mar;32(2):234-58
references.pmid: 18266856
references.citation: Nucleic Acids Res. 2007;35(6):e45
references.pmid: 17317681
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Jun;61(Pt 6):651-7
references.pmid: 15930615
references.citation: Rev Infect Dis. 1983 Mar-Apr;5 Suppl 1:S108-26
references.pmid: 6221381
references.citation: Microbiol Mol Biol Rev. 1998 Dec;62(4):1079-93
references.pmid: 9841666
references.citation: J Pharmacol Exp Ther. 1985 Nov;235(2):382-8
references.pmid: 2865352
references.citation: Clin Microbiol Rev. 2001 Oct;14(4):933-51, table of contents
references.pmid: 11585791
references.citation: Antimicrob Agents Chemother. 1979 Nov;16(5):625-30
references.pmid: 118706
references.citation: Protein Expr Purif. 2005 May;41(1):207-34
references.pmid: 15915565