Secretory protein profiling reveals TNF-α inactivation by selective and promiscuous Sec61 modulators

Cotransins are cyclic heptadepsipeptides that bind the Sec61 translocon to inhibit cotranslational translocation of a subset of secreted and type I transmembrane proteins. The few known cotransin-sensitive substrates are all targeted to the translocon by?a cleavable signal sequence, previously shown to be a critical determinant of cotransin sensitivity. By profiling two cotransin variants against a panel of secreted and transmembrane proteins, we demonstrate that cotransin side-chain differences profoundly affect substrate selectivity. Among the most sensitive substrates we identified is the proinflammatory cytokine tumor necrosis factor alpha (TNF-α). Like all type II transmembrane proteins, TNF-α is targeted to the translocon by its membrane-spanning domain, indicating that a cleavable signal sequence is not strictly required for cotransin sensitivity. Our results thus reveal an unanticipated breadth of translocon substrates whose expression is inhibited by Sec61 modulators.     

Density of states of a two-dimensional electron in a strong magnetic field and a random potential

The density of states of a two-dimensional electron in a strong magnetic field moving in a periodic and a random potential is calculated. The results are compared with the density of states of the Landau model with disorder as obtained in the single band approximation. The limitations of the single band model are discussed.     

Lithium environment in dilute poly(ethylene oxide)/lithium triflate polymer electrolyte from REDOR NMR spectroscopy

The role of the lithium ion environment is of fundamental interest regarding transport and conductivity in lithium polymer electrolytes. X-ray crystallography has been used to characterize the lithium environment in completely crystalline poly(ethylene oxide) (PEO) electrolytes, but this approach cannot be used with dilute PEO electrolytes. Here, using solid-state NMR data collected with the rotational-echo double-resonance 13C[7Li] (REDOR) pulse sequence, we have been able to characterize the crystalline microdomains of a PEO-lithium triflate sample with an oxygen/lithium ratio of 20:1. Our data clearly demonstrates that the lithium crystalline microdomains are nearly identical to those of a completely crystalline 3:1 sample, for which the crystal structure is known.     

Swedish employment offices: A new model for evaluating effectiveness

We measure how well Swedish employment offices perform in delivering the services required of them by the Swedish government. In contrast to earlier studies we use a dynamic efficiency framework, which allows us to better model the intertemporal nature of these services, explicitly allowing for placements of intermediate nature across periods. Rather than using second stage analysis to assess the effects of varying local labor market conditions and differences in client characteristics on performance, we include a measure of the office’s expected work load directly in the model. This measure, derived from duration analysis, is designed to capture the variation across offices in resources needed before an average individual can obtain employment. It is estimated from the characteristics of all unemployed individual and local labor market conditions.     

Effect of laser intensity on fast-electron-beam divergence in solid-density plasmas

Metal foil targets were irradiated with 1 mum wavelength (lambda) laser pulses of 5 ps duration and focused intensities (I) of up to 4x10;{19} W cm;{-2}, giving values of both Ilambda;{2} and pulse duration comparable to those required for fast ignition inertial fusion. The divergence of the electrons accelerated into the target was determined from spatially resolved measurements of x-ray K_{alpha} emission and from transverse probing of the plasma formed on the back of the foils. Comparison of the divergence with other published data shows that it increases with Ilambda;{2} and is independent of pulse duration. Two-dimensional particle-in-cell simulations reproduce these results, indicating that it is a fundamental property of the laser-plasma interaction.     

Improved Molecular Weight Control in Ring‐Opening Metathesis Polymerization (ROMP) Reactions with Ru‐Based Olefin Metathesis Catalysts Using N Donors and Acid: A Kinetic and Mechanistic Investigation

The effect of the addition of H₃PO₄ on the ROMP activity of cyclooctene (COE) with first‐ [Cl₂(PCy₃)₂Ru?CHPh] and second‐generation [(H₂IMes)Cl₂(PCy₃)Ru?CHPh] Grubbs’ catalysts 1 and 4 (Cy=cyclohexyl, Ph=phenyl, Mes=2,4,6‐trimethylphenyl (mesityl)), their inhibited mixtures with 1‐methylimidazole (MIM), as well as their isolated bis‐N,N′‐dimethylaminopyridine (DMAP) derivatives [Cl₂(PCy₃)(DMAP)₂Ru?CHPh)] ( 5 b ) and [Cl₂(H₂IMes)(DMAP)₂Ru?CHPh] ( 7 b ) (DMAP=dimethylaminopyridine), a novel catalyst, has been investigated. The studies include the determination of their initiation rates, as well as a determination of the molecular weights and molecular weight distributions of the polymers obtained with these catalysts and catalyst mixtures from the exo‐7‐oxanorbornene derivative 11 . The structure of catalyst 7 b was confirmed by means of X‐ray diffraction. All N‐donor‐bearing catalysts or N‐donor‐containing catalyst mixtures not only exhibited elevated activity in the presence of acid, but also increased initiation rates. Using the reversible inhibition/activation protocol with MIM and H₃PO₄ enabled us to conduct controlled ROMP with catalyst 4 producing the isolated exo‐7‐oxanorbornene‐based polymer 12 with predetermined molecular weights and narrow molecular weight distributions. This effect was based on fast and efficient catalyst initiation in contrast to the parent catalyst 4 . Hexacoordinate complex 5 b also experienced a dramatic increase in initiation rates upon acid‐addition and the ROMP reactions became well‐controlled in contrast to the acid‐free reaction. In contrast, complex 7 b performs well‐controlled ROMP in the absence of acid, whereas the polymerization of the same monomer becomes less controlled in the presence of H₃PO₄. The closer evaluation of catalysts 5 b and 7 b demonstrated that their initiation rates exhibit a linear dependency on the substrate concentration in contrast to catalysts 1 and 4 . As a consequence, their initiation rates are determined by an associative step, not a dissociative step as seen for catalysts 1 and 4 . A feasible associative metathesis initiation mechanism is proposed.     

Mobility edge in the three dimensional Anderson model

The localization length as a function of energy and disorder of a three dimensional disordered system described by the Anderson Hamiltonian is determined. The phase diagram for localization is discussed with particular emphasis on the mechanisms which are important for localization (quantum interference and tunneling).     

Positivity‐preserving,flux‐limited finite‐difference and finite‐element methods for reactive transport

A new class of positivity‐preserving, flux‐limited finite‐difference and Petrov–Galerkin (PG) finite‐element methods are devised for reactive transport problems.The methods are similar to classical TVD flux‐limited schemes with the main difference being that the flux‐limiter constraint is designed to preserve positivity for problems involving diffusion and reaction. In the finite‐element formulation, we also consider the effect of numerical quadrature in the lumped and consistent mass matrix forms on the positivity‐preserving property. Analysis of the latter scheme shows that positivity‐preserving solutions of the resulting difference equations can only be guaranteed if the flux‐limited scheme is both implicit and satisfies an additional lower‐bound condition on time‐step size. We show that this condition also applies to standard Galerkin linear finite‐element approximations to the linear diffusion equation. Numerical experiments are provided to demonstrate the behavior of the methods and confirm the theoretical conditions on time‐step size, mesh spacing, and flux limiting for transport problems with and without nonlinear reaction. Copyright © 2003 John Wiley & Sons, Ltd.     

Compact high-order finite-element method for elliptic transport problems with variable coefficients

We present a new conforming bilinear Petrov-Galerkin finite-element scheme for elliptic transport problems with variable coefficients. This scheme combines a generalized test function and artificial diffusion to achieve O(h⁴) grid-point accuracy on uniform stencils of 3 × 3 in two dimensions without resorting to the extended stencils of high-order elements. The method is compared with upwind and high-order finite-difference schemes and the standard Galerkin finite-element method for representative test problems. © 1994 John Wiley & Sons, Inc.     

Tandem mass spectrometry investigation of ADP-ribosylated kemptide

Bacterial adenosine diphosphate-ribosyltransferases (ADPRTs) are toxins that play a significant role in pathogenicity by inactivating host proteins through covalent addition of ADP-ribose. In this study we used ADP-ribosylated Kemptide (LRRASLG) as a standard to examine the effectiveness of three common tandem mass spectrometry fragmentation methods for assignment of amino acid sequence and site of modification. Fragmentation mechanisms investigated include low-energy collision-induced dissociation (CID), infrared multiphoton dissociation (IRMPD), and electron-capture dissociation (ECD); all were performed on a hybrid linear ion trap Fourier transform ion cyclotron resonance mass spectrometer. We show that ECD, but neither CID nor IRMPD, of ADP-ribosylated Kemptide produces tandem mass spectra that are interpretable with regard to amino acid sequence assignment and site of modification. Examination of CID and IRMPD tandem mass spectra of ADP-ribosylated Kemptide revealed that fragmentation was primarily focused to the ADP-ribose region, generating several potential diagnostic ions for use in discovery of ADP-ribosylated proteins. Because of the lower relative sensitivity of ECD during data-dependent acquisition to CID, we suggest a 2-fold strategy where CID and IRMPD are first used to detect ADP-ribosylated peptides, followed by sequence assignment and location of modification by ECD analysis.