Biapenem Inactivation by B2 Metallo β-Lactamases: Energy Landscape of the Post-Hydrolysis Reactions

<div><h3>Background</h3><p>The first line of defense by bacteria against <em>β</em>-lactam antibiotics is the expression of β-lactamases, which cleave the amide bond of the β-lactam ring. In the reaction of biapenem inactivation by B2 metallo β-lactamases (MβLs), after the β-lactam ring is opened, the carboxyl group generated by the hydrolytic process and the hydroxyethyl group (common to all carbapenems) rotate around the C5–C6 bond, assuming a new position that allows a proton transfer from the hydroxyethyl group to C2, and a nucleophilic attack on C3 by the oxygen atom of the same side-chain. This process leads to the formation of a bicyclic compound, as originally observed in the X-ray structure of the metallo β-lactamase CphA in complex with product.</p> <h3>Methodology/Principal Findings</h3><p>QM/MM and metadynamics simulations of the post-hydrolysis steps in solution and in the enzyme reveal that while the rotation of the hydroxyethyl group can occur in solution or in the enzyme active site, formation of the bicyclic compound occurs primarily in solution, after which the final product binds back to the enzyme. The calculations also suggest that the rotation and cyclization steps can occur at a rate comparable to that observed experimentally for the enzymatic inactivation of biapenem only if the hydrolysis reaction leaves the N4 nitrogen of the β-lactam ring unprotonated.</p> <h3>Conclusions/Significance</h3><p>The calculations support the existence of a common mechanism (in which ionized N4 is the leaving group) for carbapenems hydrolysis in all MβLs, and suggest a possible revision of mechanisms for B2 MβLs in which the cleavage of the β-lactam ring is associated with or immediately followed by protonation of N4. The study also indicates that the bicyclic derivative of biapenem has significant affinity for B2 MβLs, and that it may be possible to obtain clinically effective inhibitors of these enzymes by modification of this lead compound.</p> </div

Dynamic mechanisms of He single ionization by fast proton impact

Triple-differential ionization cross sections 3 pdcphipdprec, the momentum distributions of singly charged recoil ions transverse to the beam direction as a function of the projectile polar (p), and azimuthal (cphip) scattering angle were measured in order to elucidate the dynamics of 3-MeV H+ on He single ionization. For projectile polar deflections 0.2 p1 mrad and azimuthal scattering angles 0°cphip360°, the kinematic regimes where two-body interactions dominate the three-body momentum exchange of the single-ionization reaction were separated experimentally. © 1992 The American Physical Society

A semiempirical molecular orbital and dynamic NMR study of conformational isomerism in angiotensin-converting enzyme inhibitors

The conformational isomerism in a series of molecules related to inhibitors of the angiotensinconverting enzyme (ACE) is investigated using the semiempirical quantum mechanical molecular orbital model, AM1, and dynamic NMR spectroscopy. The theoretical method is tested by the prediction of relative barriers to rotation about amide C-N and acylhydrazine N-N bonds, and of the trans: cis ratio for a series of model compounds. The conformational energetics of captopril and a series of acylhydrazines are predicted and used to interpret the dynamic NMR spectra of these molecules, presented herein. © 1991

An activated sulfonylating agent that undergoes general base-catalyzed hydrolysis by amines in preference to aminolysis

Activated sulfonyl derivatives, similar to acyl ones, usually undergo aminolysis with amines in water as nucleophilic attack by the amine is preferred to hydrolysis. However, despite being active sulfonyl derivatives, four-membered heterocyclic sulfonamides, β-sultams, do not undergo aminolysis in aqueous solution but preferentially react to give hydrolysis products only. The rate of the reaction of β-sultams in buffered solutions of simple primary amines shows a first-order dependence on amine concentrations attributed to general base-catalyzed hydrolysis by the amine. Even N-benzyl-4,4-dimethyl-3-oxo-β-sultam, which is both a β-sultam and a β-lactam, undergoes hydrolysis at the sulfonyl center rather than aminolysis at either the sulfonyl or acyl center. The solvent kinetic isotope effects (SKIE, kH2O/kD2O) for the amine-catalyzed hydrolyses are 1.4 and 1.9 for the hydrolysis of N-benzoyl-β-sultam and N-benzyl-4,4-dimethyl-3-oxo-β-sultam, respectively, compatible with a general base-catalyzed mechanism. The amine-catalyzed hydrolysis gives a Bronsted β value of +0.9 for both N-benzoyl β-sultam and N-benzyl-4,4-dimethyl-3-oxo-β-sultam, indicating that the general base amine is almost fully protonated in the transition state. A general base-catalyzed mechanism for hydrolysis rather than nucleophilic attack was also deduced for the reaction of N-benzyl-4,4-dimethyl-3-oxo-β-sultam with carboxylate anions based on a SKIE of 1.7−1.9 and rate constants which fit the Bronsted plot for amines. In contrast to acyl transfer reactions, those for sulfonyl transfer appear to show an inverse reactivity-selectivity relationshipthe most active compounds being the most selective. The lack of reactivity of β-sultams toward amine nucleophiles appears to be related to the mechanism of ring opening of β-sultams with a decreased reactivity toward amines relative to hydroxide ion, probably related to the expulsion of the relatively poor leaving group amide anion