5mmd

TMB-1. Structural insights into TMB-1 and the role of residue 119 and 228 in substrate and inhibitor binding

spinqualitylabelsall models 5mmd, resolution 1.75Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 5mmd is a 2 chain structure with sequence from Acinetobacter baumannii. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: X-ray diffraction, Resolution 1.75Å Ligands: CL, ZN Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function T2HNV0_ACIBA Publication Abstract from PubMed Metallo-beta-lactamases (MBLs) threaten the effectiveness of beta-lactam antibiotics, including carbapenems, and are a concern for global public health. beta-Lactam/beta-lactamase inhibitor combinations active against class A and class D carbapenemases are used, but no clinically useful MBL inhibitor is currently available. Tripoli metallo-beta-lactamase-1 (TMB-1) and TMB-2 are members of MBL subclass B1a, where TMB-2 is an S228P variant of TMB-1. The role of S228P was studied by comparisons of TMB-1 and TMB-2, and E119 was investigated through the construction of site-directed mutants of TMB-1, E119Q, E119S, and E119A (E119Q/S/A). All TMB variants were characterized through enzyme kinetic studies. Thermostability and crystallization analyses of TMB-1 were performed. Thiol-based inhibitors were investigated by determining the 50% inhibitory concentrations (IC50) and binding using surface plasmon resonance (SPR) for analysis of TMB-1. Thermostability measurements found TMB-1 to be stabilized by high NaCl concentrations. Steady-state enzyme kinetics analyses found substitutions of E119, in particular, substitutions associated with the penicillins, to affect hydrolysis to some extent. TMB-2 with S228P showed slightly reduced catalytic efficiency compared to TMB-1. The IC50 levels of the new thiol-based inhibitors were 0.66 muM (inhibitor 2a) and 0.62 muM (inhibitor 2b), and the equilibrium dissociation constant (KD ) of inhibitor 2a was 1.6 muM; thus, both were more potent inhibitors than l-captopril (IC50 = 47 muM; KD = 25 muM). The crystal structure of TMB-1 was resolved to 1.75 A. Modeling of inhibitor 2b in the TMB-1 active site suggested that the presence of the W64 residue results in T-shaped pi-pi stacking and R224 cation-pi interactions with the phenyl ring of the inhibitor. In sum, the results suggest that residues 119 and 228 affect the catalytic efficiency of TMB-1 and that inhibitors 2a and 2b are more potent inhibitors for TMB-1 than l-captopril. Structural Insights into TMB-1 and the Role of Residues 119 and 228 in Substrate and Inhibitor Binding.,Skagseth S, Christopeit T, Akhter S, Bayer A, Samuelsen O, Leiros HS Antimicrob Agents Chemother. 2017 Jul 25;61(8). pii: e02602-16. doi:, 10.1128/AAC.02602-16. Print 2017 Aug. PMID:28559248 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Beta-lactamase 3D structures

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