title: Structure and inhibition of N-acetylneuraminate lyase from methicillin-resistant Staphylococcus aureus.
abstract: N-Acetylneuraminate lyase is the first committed enzyme in the degradation of sialic acid by bacterial pathogens. In this study, we analyzed the kinetic parameters of N-acetylneuraminate lyase from methicillin-resistant Staphylococcus aureus (MRSA). We determined that the enzyme has a relatively high KMof 3.2 mm, suggesting that flux through the catabolic pathway is likely to be controlled by this enzyme. Our data indicate that sialic acid alditol, a known inhibitor of N-acetylneuraminate lyase enzymes, is a stronger inhibitor of MRSA N-acetylneuraminate lyase than of Clostridium perfringens N-acetylneuraminate lyase. Our analysis of the crystal structure of ligand-free and 2R-sialic acid alditol-bound MRSA N-acetylneuraminate lyase suggests that subtle dynamic differences in solution and/or altered binding interactions within the active site may account for species-specific inhibition.
authors.affiliation: Biomolecular Interaction Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: North
authors.affiliation: Department of Chemistry, University of Canterbury, Christchurch, New Zealand.
authors.forename: Andrew J A
authors.initials: AJ
authors.lastname: Watson
authors.affiliation: Biomolecular Interaction Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.
authors.forename: F Grant
authors.initials: FG
authors.lastname: Pearce
authors.affiliation: Protein Science and Engineering, Callaghan Innovation, University of Canterbury, Christchurch, New Zealand.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Muscroft-Taylor
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Sweden.|Centre for Antibiotic Resistance Research (CARe), University of Gothenburg, Sweden.|Department of Structural Biology, School of Medicine, Stanford University, CA, USA.
authors.forename: Rosmarie
authors.initials: R
authors.lastname: Friemann
authors.affiliation: Biomolecular Interaction Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.|Department of Chemistry, University of Canterbury, Christchurch, New Zealand.
authors.forename: Antony J
authors.initials: AJ
authors.lastname: Fairbanks
authors.affiliation: Biomolecular Interaction Centre, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.|Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Australia.
authors.forename: Renwick C J
authors.identifier: http://orcid.org/0000-0002-5506-4939
authors.initials: RC
authors.lastname: Dobson
mesh_terms: D000595:Amino Acid Sequence; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D055624:Methicillin-Resistant Staphylococcus aureus / Q000201:enzymology*; D008958:Models, Molecular; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D007652:Oxo-Acid-Lyases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D020836:Protein Structure, Quaternary; D013045:Species Specificity
chemical_list: D004791:Enzyme Inhibitors; D007652:Oxo-Acid-Lyases; C018728:N-acetylneuraminate lyase; D019158:N-Acetylneuraminic Acid
keywords: N-acetylneuraminate lyase; antibiotic resistance; drug discovery; inhibitor; methicillin-resistant Staphylococcus aureus; sialic acid degradation; structure