Bacterial Hsp70 resolves misfolded states and accelerates productive folding of a multi-domain protein

The ATP-dependent Hsp70 chaperones (DnaK in E. coli) mediate protein folding in cooperation with J proteins and nucleotide exchange factors (E. coli DnaJ and GrpE, respectively). The Hsp70 system prevents protein aggregation and increases folding yields. Whether it also enhances the rate of folding remains unclear. Here we show that DnaK/DnaJ/GrpE accelerate the folding of the multi-domain protein firefly luciferase (FLuc) 20-fold over the rate of spontaneous folding measured in the absence of aggregation. Analysis by single-pair FRET and hydrogen/deuterium exchange identified inter-domain misfolding as the cause of slow folding. DnaK binding expands the misfolded region and thereby resolves the kinetically-trapped intermediates, with folding occurring upon GrpE-mediated release. In each round of release DnaK commits a fraction of FLuc to fast folding, circumventing misfolding. We suggest that by resolving misfolding and accelerating productive folding, the bacterial Hsp70 system can maintain proteins in their native states under otherwise denaturing stress conditions. The Hsp70 system prevents protein aggregation and increases folding yields, but it is unknown whether it also enhances the rate of folding. Here the authors combine refolding assays, FRET and hydrogen/deuterium exchange-mass spectrometry measurements to study the folding of firefly luciferase and find that the bacterial Hsp70 actively promotes the folding of this multi-domain protein

Quadratic maps between modules

We introduce a notion of $R$-quadratic maps between modules over a commutative ring $R$ which generalizes several classical notions arising in linear algebra and group theory. On a given module $M$ such maps are represented by $R$-linear maps on a certain module $P^2_R(M)$. The structure of this module is described in term of the symmetric tensor square $Sym^2_R(M)$, the degree 2 component $\Gamma^2_R(M)$ of the divided power algebra over $M$, and the ideal $I_2$ of $R$ generated by the elements $r^2-r$, $r\in R$. The latter is shown to represent quadratic derivations on $R$ which arise in the theory of modules over square rings. This allows to extend the classical notion of nilpotent $R$-group of class 2 with coefficients in a 2-binomial ring $R$ to any ring $R$. We provide a functorial presentation of $I_2$ and several exact sequences embedding the modules $P^2_R(M)$ and $\Gamma^2_R(M)$.Comment: 22 page

On the Determination of the Polarized Sea Distributions of the Nucleon

The possibilities to determine the flavor structure of the polarized sea (antiquark) distributions of the nucleon via vector boson $(\gamma^*, W^{\pm}, Z^0)$ production at high energy polarized hadron--hadron colliders, such as the Relativistic Heavy--Ion Collider (RHIC), are studied in detail. In particular the perturbative stability of the expected asymmetries in two representative models for the (un)broken flavor structure are investigated by confronting perturbative QCD leading order predictions of the expected asymmetries with their next--to--leading order counterparts.Comment: 28 pages, LaTe

The proteostasis network and its decline in ageing

Ageing is a major risk factor for the development of many diseases, prominently including neurodegenerative disorders such as Alzheimer disease and Parkinson disease. A hallmark of many age-related diseases is the dysfunction in protein homeostasis (proteostasis), leading to the accumulation of protein aggregates. In healthy cells, a complex proteostasis network, comprising molecular chaperones and proteolytic machineries and their regulators, operates to ensure the maintenance of proteostasis. These factors coordinate protein synthesis with polypeptide folding, the conservation of protein conformation and protein degradation. However, sustaining proteome balance is a challenging task in the face of various external and endogenous stresses that accumulate during ageing. These stresses lead to the decline of proteostasis network capacity and proteome integrity. The resulting accumulation of misfolded and aggregated proteins affects, in particular, postmitotic cell types such as neurons, manifesting in disease. Recent analyses of proteome-wide changes that occur during ageing inform strategies to improve proteostasis. The possibilities of pharmacological augmentation of the capacity of proteostasis networks hold great promise for delaying the onset of age-related pathologies associated with proteome deterioration and for extending healthspan

Local P-shtukas and their relation to global G-shtukas

This is the first in a sequence of two articles investigating moduli stacks of global G-shtukas, which are function field analogs for Shimura varieties. Here G is a flat affine group scheme of finite type over a smooth projective curve, and global G-shtukas are generalizations of Drinfeld shtukas and analogs of abelian varieties with additional structure. Our moduli stacks generalize various moduli spaces used by different authors to prove instances of the Langlands program over function fields. In the present article we explain the relation between global G-shtukas and local P-shtukas, which are the function field analogs of p-divisible groups with additional structure. We prove the analog of a theorem of Serre and Tate stating the equivalence between the deformations of a global G-shtuka and its associated local P-shtukas. We also investigate local P-shtukas alone and explain their relation with Galois representations through their Tate modules. And if P is a smooth affine group scheme with connected reductive generic fiber we prove the existence of Rapoport--Zink spaces for bounded local P-shtukas as formal schemes locally formally of finite type. In the sequel to this article we use these Rapoport--Zink spaces to uniformize the moduli stacks of global G-shtukas.Comment: 37 pages, v3: generalization to flat affine group schemes of finite type, v3: final version which appears in Muenster J. of Mathematic

The Deterministic Impulse Control Maximum Principle in Operations Research: Necessary and Sufficient Optimality Conditions (replaces CentER DP 2011-052)

This paper considers a class of optimal control problems that allows jumps in the state variable. We present the necessary optimality conditions of the Impulse Control Maximum Principle based on the current value formulation. By reviewing the existing impulse control models in the literature, we point out that meaningful problems do not satisfy the sufficiency conditions. In particular, such problems either have a concave cost function, contain a fixed cost, or have a control-state interaction, which have in common that they each violate the concavity hypotheses used in the sufficiency theorem. The implication is that the corresponding problem in principle has multiple solutions that satisfy the necessary optimality conditions. Moreover, we argue that problems with fixed cost do not satisfy the conditions under which the necessary optimality conditions can be applied. However, we design a transformation, which ensures that the application of the Impulse Control Maximum Principle still provides the optimal solution. Finally, we show for the first time that for some existing models in the literature no optimal solution exists.Impulse Control Maximum Principle;Optimal Control;discrete continuous system;state-jumps;present value formulation.

Novel Method to Process Cystic Fibrosis Sputum for Determination of Oxidative State

Background: Induced sputum is the most commonly used method to analyze airway inflammation in cystic fibrosis (CF) patients ex vivo. Due to the complex matrix of the sample material, precise and reliable analysis of sputum constituents depends critically on preanalytical issues. Objectives: Here we compared the commonly used method for sputum processing by dithiothreitol (DTT) with a novel mechanical method in regard to basal cellular parameters, neutrophil markers and glutathione (GSH) levels. Methods: Sputum samples from CF patients were processed in parallel with or without the use of DTT. The key improvement of the mechanical method was the processing in many very small aliquots. Cellular and humoral markers were assessed and compared according to Bland-Altman. Results: Total cell count, cell viability, differential cell count, neutrophil elastase levels and flow cytometrically analyzed neutrophil markers (CD63, CD11b, DHR) did not differ between the two methods. Intracellular and extracellular GSH levels were significantly higher in DTT-treated samples (p = 0.002). Conclusion: The mechanical sputum-processing method presented had a similar yield of cells and fluids as the conventional DTT method and the advantage of omitting the introduction of reducing agents. This method allows a more reliable analysis of redox-dependent airway inflammation in sputum cells and fluid from CF patients than methods utilizing DTT. Copyright (C) 2009 S. Karger AG, Base

Genotype and environment interaction on yield and quality parameters of organically grown winter wheat – Triticum aestivum L. genotypes

The interaction of genotype and environment upon yield and quality parameters of eight winter wheat (Triticum aestivum L.) genotypes was studied under organic conditions in Austria over two growing periods, 2001/2002 and 2002/2003, respectively. Two sites that have significantly different climatic conditions, Innviertel and Marchfeld, were chosen for the field experiment. Study site weather and soil conditions are important yield-affecting factors. Although the yield of Marchfeld-grown genotypes were lower, they had shown higher quality parameter values. Soil moisture conditions increase the grain yield but decrease its quality. To obtain seed with higher quality, a production site with favourable climate conditions should be chosen