Antibiotic recognition by binuclear metallo-beta-lactamases revealed by X-ray crystallography

Metallo-beta-lactamases are zinc-dependent enzymes responsible for resistance to beta-lactam antibiotics in a variety of host bacteria, usually Gram-negative species that act as opportunist pathogens. They hydrolyze all classes of beta-lactam antibiotics, including carbapenems, and escape the action of available beta-lactamase inhibitors. Efforts to develop effective inhibitors have been hampered by the lack of structural information regarding how these enzymes recognize and turn over beta-lactam substrates. We report here the crystal structure of the Stenotrophomonas maltophilia L1 enzyme in complex with the hydrolysis product of the 7alpha-methoxyoxacephem, moxalactam. The on-enzyme complex is a 3'-exo-methylene species generated by elimination of the 1-methyltetrazolyl-5-thiolate anion from the 3'-methyl group. Moxalactam binding to L1 involves direct interaction of the two active site zinc ions with the beta-lactam amide and C4 carboxylate, groups that are common to all beta-lactam substrates. The 7beta-[(4-hydroxyphenyl)malonyl]-amino substituent makes limited hydrophobic and hydrogen bonding contacts with the active site groove. The mode of binding provides strong evidence that a water molecule situated between the two metal ions is the most likely nucleophile in the hydrolytic reaction. These data suggest a reaction mechanism for metallo-beta-lactamases in which both metal ions contribute to catalysis by activating the bridging water/hydroxide nucleophile, polarizing the substrate amide bond for attack and stabilizing anionic nitrogen intermediates. The structure illustrates how a binuclear zinc site confers upon metallo-beta-lactamases the ability both to recognize and efficiently hydrolyze a wide variety of beta-lactam substrates.     

On the flexibility of the design of multiple try Metropolis schemes

The multiple try Metropolis (MTM) method is a generalization of the classical Metropolis–Hastings algorithm in which the next state of the chain is chosen among a set of samples, according to normalized weights. In the literature, several extensions have been proposed. In this work, we show and remark upon the flexibility of the design of MTM-type methods, fulfilling the detailed balance condition. We discuss several possibilities, show different numerical simulations and discuss the implications of the results.     

Effectiveness of stirring measures in an axisymmetric rotating annulus flow

Lagrangian stirring in a thermally driven rotating annulus is investigated numerically using a Navier-Stokes model and a second order Runge-Kutta integration routine. The stirring properties are investigated using finite scale Lyapunov exponents, Lagrangian coherent structures and a leaking method. The ability of these measures to identify transport barriers, regions of well and poorly stirred flow, and stable and unstable manifolds is investigated, as well as the stirring properties of the annulus flow. It is found that finite scale Lyapunov exponents characterise the stirring properties of flows occurring in the rotating annulus more efficiently than the leaking method or Lagrangian coherent structures. The strength of the stirring varies monotonically with thermal forcing amplitude, but non-monotonically with forcing frequency. The flows investigated are axisymmetric (i.e. two dimensional) and time dependent.     

Cell-penetrant, nanomolar O-GlcNAcase inhibitors selective against lysosomal hexosaminidases

Posttranslational modification of metazoan nucleocytoplasmic proteins with N-acetylglucosamine (O-GlcNAc) is essential, dynamic, and inducible and can compete with protein phosphorylation in signal transduction. Inhibitors of O-GlcNAcase, the enzyme removing O-GlcNAc, are useful tools for studying the role of O-GlcNAc in a range of cellular processes. We report the discovery of nanomolar OGA inhibitors that are up to 900,000-fold selective over the related lysosomal hexosaminidases. When applied at nanomolar concentrations on live cells, these cell-penetrant molecules shift the O-GlcNAc equilibrium toward hyper-O-GlcNAcylation with EC?? values down to 3 nM and are thus invaluable tools for the study of O-GlcNAc cell biology.     

C*-Algebras with Dense Nilpotent Subalgebras

In a beautiful result, Herrero (D. A. Herrero, ‘Normallimits of nilpotent operators’, Indiana Univ. Math. J.23 (1973/74) 1097–1108) showed that a normal operatoron l² lies in the closure of the set of nilpotent operatorsif and only if its spectrum is connected and contains zero.In the quest for an automatic continuity result for algebrahomomorphisms between C* -algebras, Dales showed that, if adiscontinuous algebra homomorphism : A u exists between C*-algebrasA and u, and if (A) is dense in u, then there is a C*-algebrau₂ with a dense subalgebra N u² such that every x N is quasinilpotent(see p. 685 of H. G. Dales, Banach algebras and automatic continuity,London Mathematical Society Monographs 24, Oxford UniversityPress, 2001). (A discontinuous homomorphism ₂: A₂ u₂ can bedefined with the same basic properties as , but the revisedtarget space u₂ has a dense subalgebra consisting of quasinilpotentelements.) As remarked by Dales, no such C*-algebra was thenknown; but here we present one. Indeed, using the full powerof Herrero's result, one may arrange that every x N is nilpotent.The C*-algebra is constructed in a ‘neat’ way; itis most naturally constructed as a non-separable, concrete C*-algebraof operators on a separable Hilbert space K but one can arrangethat the algebra u itself be separable if desired. 2000 MathematicsSubject Classification 47C15, 46H40 (primary), 47A10, 46L06,46L05, 46H35 (secondary).