title: Structural insight into mode of binding of Meropenem to CTX-M-15 type β-lactamase.
abstract: Among Enterobacteriaceae, CTX-M type extended spectrum beta lactamase confers potent hydrolytic activity against cephalosporin group of antibiotics. Strains producing CTX-M type beta lactamase enzymes, show high level of resistance against cefotaxime. Therefore carbapenem antibiotics are used against beta lactamase producing strains. Hence, this study was designed to understand an insight of molecular basis of CTX-M-15 interaction with meropenem, and its effect on CTX-M-15 efficiency. Clinical strain of Enterobacter cloacae (EC-15) was used to clone blaCTX-M-15gene in E.coli BL21cells. The protein was then expressed and purified. Results showed that CTX-M-15 producing strains are susceptible to meropenem. It quenches the fluorescence of CTX-M-15 spontaneously with binding constant of the order of 103M-1. Meropenem binds on the active site of CTX-M-15, hydrogen bonded with four common amino acid residues of cefotaxime binding site, as revealed by molecular docking studies. Conformational change in the structure of CTX-M-15 was observed upon meropenem binding by CD spectroscopy. The catalytic efficiency of CTX-M-15 was decreased up to 4 times upon meropenem binding. Docking study shows that few amino acids of active site of enzyme are also involved in meropenem binding, hence substrate is difficult to bind on active site properly and does not get hydrolysed. Moreover, meropenem binding induces structural changes in CTX-M-15, making the enzyme less efficient.
authors.affiliation: Medical Microbiology and Molecular Biology Laboratory, Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, Uttar Pradesh 202 002, India.
authors.forename: Lubna
authors.initials: L
authors.lastname: Maryam
authors.affiliation: Medical Microbiology and Molecular Biology Laboratory, Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, Uttar Pradesh 202 002, India. Electronic address: asad.k@rediffmail.com.
authors.forename: Asad U
authors.initials: AU
authors.lastname: Khan
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism*; D016972:Enterobacter cloacae / Q000201:enzymology; D006860:Hydrogen Bonding; D007700:Kinetics; D000077731:Meropenem; D062105:Molecular Docking Simulation; D011485:Protein Binding; D011487:Protein Conformation; D013845:Thienamycins / Q000737:chemistry* / Q000378:metabolism*; D001618:beta-Lactamases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D013845:Thienamycins; C473109:beta-lactamase CTX-M-15; D001618:beta-Lactamases; D000077731:Meropenem
keywords: Antibiotic resistance; Beta-lactamase; Fluorescence quenching; Meropenem; Molecular docking