Quorum-quenching proteins as therapeutic targets: Binding and reaction mechanism

COMP 401

Katrin Spiegel, spiegel@cmm.upenn.edu1, Marco De Vivo, mdevivo@cmm.upenn.edu2, Matteo Dal Peraro, matteodp@cmm.upenn.edu2, and Michael L. Klein, klein@seas.upenn.edu1. (1) Department of Chemistry, University of Pennsylvania, 231 S. 34th Street, Phialdelphia, PA 19104-6323, (2) Center for Molecular Modeling, Chemistry Department, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104
N-acyl-L-homoserine lactones (AHL's) are believed to be involved in cell-to-cell communication, regulating gene expression and group behavior in bacteria. This phenomenon is also referred to as quorum sensing (QS); thus, quorum-quenching proteins, such as AHL-Lactonases constitute an appealing new target for therapeutic approaches to control pathogens. Based on the recent crystal structure of the inhibitor-enzyme complex of AHL-lactonase of Bacillus thuringiensis at 1.7 Å (1), we apply an extensive multi-step computational investigation with the scope to rationalize the structure-activity data for real substrates and, hence, better understand the mechanism of AHL-Lactonases. A standard docking procedure is applied to identify possible substrate binding modes followed by the modeling of a mobile loop region, not defined in the experimental structure. Then, classical molecular dynamics simulations have been performed in order to further refine the best docked structures and to equilibrate the solvated system. Finally, we discriminate among possible binding modes by looking at the enzymatic reaction energy profiles and comparing them to experimental data. To this aim, Car-Parrinello QM/MM simulations together with an Umbrella-sampling scheme are performed. We confront our findings to the reaction mechanism of other members of the metallo-β-lactamase folding family (e.g. CcrA from Bacteroides fragilis, and Glyoxalase II).

(1) Kim M.H., Choi W.-C., Kang H.O., Lee J.S., Kang B.S., Kim K.-J., Derewenda Z.S., Oh T.-K., Lee C.H., Lee J.-K. PNAS, 102(49):17606-17611 (2005).