Crystal structure of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26,

ABSTRACT: The haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB) is the enzyme involved in the degradation of the important environmental pollutant γ-hexachlorocyclohexane. The enzyme hydrolyzes a broad range of halogenated cyclic and aliphatic compounds. Here, we present the 1.58 Å crystal structure of LinB and the 2.0 Å structure of LinB with 1,3-propanediol, a product of debromination of 1,3-dibromopropane, in the active site of the enzyme. The enzyme belongs to the R/ hydrolase family and contains a catalytic triad (Asp108, His272, and Glu132) in the lipase-like topological arrangement previously proposed from mutagenesis experiments. The LinB structure was compared with the structures of haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 and from Rhodococcus sp. and the structural features involved in the adaptation toward xenobiotic substrates were identified. The arrangement and composition of the R-helices in the cap domain results in the differences in the size and shape of the active-site cavity and the entrance tunnel. This is the major determinant of the substrate specificity of this haloalkane dehalogenase

Iripin-1, a new anti-inflammatory tick serpin, inhibits leukocyte recruitment in vivo while altering the levels of chemokines and adhesion molecules

Serpins are widely distributed and functionally diverse inhibitors of serine proteases. Ticks secrete serpins with anti-coagulation, anti-inflammatory, and immunomodulatory activities via their saliva into the feeding cavity to modulate host’s hemostatic and immune reaction initiated by the insertion of tick’s mouthparts into skin. The suppression of the host’s immune response not only allows ticks to feed on a host for several days but also creates favorable conditions for the transmission of tick-borne pathogens. Herein we present the functional and structural characterization of Iripin-1 (Ixodes ricinus serpin-1), whose expression was detected in the salivary glands of the tick Ixodes ricinus, a European vector of tick-borne encephalitis and Lyme disease. Of 16 selected serine proteases, Iripin-1 inhibited primarily trypsin and further exhibited weaker inhibitory activity against kallikrein, matriptase, and plasmin. In the mouse model of acute peritonitis, Iripin-1 enhanced the production of the anti-inflammatory cytokine IL-10 and chemokines involved in neutrophil and monocyte recruitment, including MCP-1/CCL2, a potent histamine-releasing factor. Despite increased chemokine levels, the migration of neutrophils and monocytes to inflamed peritoneal cavities was significantly attenuated following Iripin-1 administration. Based on the results of in vitro experiments, immune cell recruitment might be inhibited due to Iripin-1-mediated reduction of the expression of chemokine receptors in neutrophils and adhesion molecules in endothelial cells. Decreased activity of serine proteases in the presence of Iripin-1 could further impede cell migration to the site of inflammation. Finally, we determined the tertiary structure of native Iripin-1 at 2.10 Å resolution by employing the X-ray crystallography technique. In conclusion, our data indicate that Iripin-1 facilitates I. ricinus feeding by attenuating the host’s inflammatory response at the tick attachment site

The impact of the crisis on the economy of Spain

This bachelor thesis describes the impact of the current economic crisis on the Spanish kingdom, the fourth largest economy of Europe. The first chapter focuses on origin and development of crisis in Spain as a whole. The attention is given to the macroeconomic indicators such as gross domestic product, unemployment, growth of the price level and public debt. In the following chapters are being compared the impacts of the crisis on two different autonomous communities of Spain, which one of them (Castile and León) is situated on the north of the country, while the other one (Andalusia) is on the south of Spain. The aim of this work is to point out the unfavourable influence of the crisis not only on the country as a whole but to compare the impacts on particular regions of Spain as well

Zefektivnění on-line komunikačního mixu oděvní společnosti JARA s.r.o.

Tato diplomová práce má za cíl navrhnout zefektivnění on-line komunikační strategie pro oděvní společnost JARA s.r.o. Společnost dosud dávala důraz na osobní doporučení zákazníků a plně nevyužívala velký potenciál digitálního marketingu. Práce je rozdělena do 3 kapitol, a to jedné teoretické a dvou praktických částí. První kapitola vysvětluje pojem on-line marketing, jeho výhody a zařazení do komunikačního mixu. Ve druhé kapitole je popsána společnost JARA s.r.o., provedena SWOT analýza, konkurenční analýza a popsána aktuální on-line komunikace společnosti, kterou je jen jednoduchá webová stránka. Třetí kapitolu tvoří provedení a výsledky terénního dotazníkového šetření uskutečněno metodou PAPI osobně na prodejně. Na základě primárního a sekundárního výzkumu je poslední podkapitolou práce návrh zkvalitnění on-line komunikačního mixu společnosti.Táto diplomová práca má za cieľ navrhnúť zefektívnenie on-line komunikačnej stratégie pre odevnú spoločnosť JARA s.r.o. Spoločnosť doteraz dávala dôraz na osobné odporúčania zákazníkov a plne nevyužívala veľký potenciál digitálneho marketingu. Práca je rozdelená do 3 kapitol, a to jednej teoretickej a dvoch praktických častí. Prvá kapitola vysvetľuje pojem on-line marketing, jeho výhody a zaradenie do komunikačného mixu. V druhej kapitole je popísaná spoločnosť JARA s.r.o., vykonaná SWOT analýza, konkurenčná analýza a popísaná aktuálna on-line komunikácia, ktorou je len jednoduchá webová stránka spoločnosti. Tretiu kapitolu tvorí prevedenie a výsledky terénneho dotazníkového šetrenia vykonané metódou PAPI osobne na predajni. Na základe primárneho a sekundárneho výskumu je poslednou podkapitolou práce návrh skvalitnenia on-line komunikačného mixu spoločnosti.This diploma thesis aims to propose an improvement in online communication strategy for company JARA ltd. This company has so far put emphasis on personal customer recommendations and does not fully exploit the great potential of digital marketing. The thesis is divided into 3 chapters: one theoretical and two practical parts. The first chapter explains the concept of on-line marketing, its benefits and its classification within the communication mix. The second chapter describes the company JARA ltd. Moreover, there are SWOT analysis and competition analysis carried out plus current online communication of the company is described, which consists of an easy company website only. The third chapter represents process and results of a field survey conducted via PAPI method personally in the stores. On the basis of primary and secondary research, the last subchapter of the thesis consists of the proposal of improvement of the company's online communication mix

WrbA bridges bacterial flavodoxins and eukaryotic NAD(P)H:quinone oxidoreductases

The crystal structure of the flavodoxin-like protein WrbA with oxidized FMN bound reveals a close relationship to mammalian NAD(P)H:quinone oxidoreductase, Nqo1. Structural comparison of WrbA, flavodoxin, and Nqo1 indicates how the twisted open-sheet fold of flavodoxins is elaborated to form multimers that extend catalytic function from one-electron transfer between protein partners using FMN to two-electron reduction of xenobiotics using FAD. The structure suggests a novel physiological role for WrbA and Nqo1

Crystallization and preliminary diffraction analysis of Escherichia coli WrbA in complex with its cofactor flavin mononucleotide

E. coli WrbA, the founding member of a novel flavoprotein family, was crystallized in complex with its physiological cofactor. Preliminary diffraction analysis is reported