935 resultados para investigation of head-first binding of substrate with the phe557 mutant soybean lipoxygenase-1


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The present paper explores, theoretically, and empirically, whether compliance with the International Code of marketing of breast-milk substitutes impacts on financial performance measured by stock markets. The empirical analysis, which considers a 20-year period, shows that stock markets are indifferent to the level of compliance by manufacturers with the International Code. Two important issues emerge from this result. Based on our finding that financial performance as measured by stock markets cannot explain the level of compliance, the first issue refers to what alternative types of mechanisms drive manufacturers who comply the least with voluntary codes such as the International Code. Conversely, from our finding that stock markets do not reward the most compliant, the second issue raised is an inherent weakness of stock markets to fully incorporate social and environmental values.

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Arboreal and terrestrial habitats impose different constraints on tetrapod locomotion. We studied Polychrus acutirostris, a tree-dwelling lizard that also moves on the ground, in order to evaluate the effects of support incline and diameter on locomotion parameters. Limb movements of six specimens were filmed to quantify kinematic variables (velocity, stride frequency, stride length, and limb coordination) on distinct perch diameters (4.0, 1.5, 0.8 cm) and inclines (90, 45, and on level ground). The results show a notable slowness in arboreal habitat combined with a relatively fast locomotion when using the ground as temporary habitat. These animals developed walking trots mainly using lateral sequence. Non-symmetrical trots adopted at the highest velocities on the ground indicate difficulties of ""accommodation"" to the constraints imposed by this condition. Velocity generally decreases with the decreasing diameter, and with increasing incline, of the supports. Slowness, gaits favouring the body stability, elective role of the stride frequency in the modulation of the speed, and the role of the hindlimb in the force exchange to propel the body, constitute the main features of the locomotion pattern of P. acutirostris.

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The objective of this study was to evaluate the chemical alterations of the substrate in the cultivation of peppers grown in coconut husk fiber, in a protected environment. Initially, 160 pepper plants ('Eppo') were divided into four blocks, where two pots per block were analyzed every 21 days after transplanting. The cultivation of pepper was carried out in plastic pots of 13 L, containing coconut husk fiber, and placed in double rows with a spacing of 0.5×0.8 m between single rows and 1.10 m between double rows. After removal of the plants from the pots, individual samples of substrate (approximately 1 L) were collected from each pot and dried at ambient temperature. The levels of Ca, Mg, S, Cl, Na, B, Fe, Mn, Cu and Zn were determined for all periods of the cultivation. These analyses were performed using the method of extraction 1:1.5 v/v. There was an increase in the levels of all the nutrients evaluated. Further studies should be conducted to develop a better nutrient solution.

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Soybean lipoxygenase-1 (SBLO-1) catalyzes the oxygenation of polyunsaturated fatty acids into conjugated diene hydroperoxides. The three dimensional structure of SBLO-1 is known, but it is not certain how substrates bind. One hypothesis involves the transient separation of helix-2 and helix-11 located on the exterior of the molecule in front of the active site iron. A second hypothesis involves a conformational change in the side chains of residues leucine 541 and threonine 259. To test these hypotheses, site directed mutagenesis was used to create a cysteine mutation on each helix, which could allow for the formation of a disulfide linkage. Disulfide formation between the two cysteines in the T259C,S545C mutant was found to be unfavorable, but later shown to be present at higher pH values using SDS-PAGE. Treatment of the T259C,S545C with the crosslinker 2,3-dibromomaleimide (DBM) resulted in a 50% reduction in catalytic activity. No loss of activity was observed when the single mutant, S545C, or the wild type was treated with DBM. Single mutants T259C and L541C both showed approximately 20% reduction in the rate after addition of DBM. Double mutants T259C,L541C and S263C,S545C showed approximately 30% reduction in the rate after addition of DBM. Single mutants T259C and L541C showed an increase in activity after incubation with NEM. Double mutants T259C,S545C and T259C,L541C showed an increase in activity after incubation with NEM. The S263C,S545C double mutant showed a slight decrease in activity in the presence of NEM. It is unclear how the NEM and DBM are interacting with the molecule, but this can easily be determined through mass spectrometry experiments.

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Moe's scale of hardness as used in mineralogy is admittedly rather indefinite, and no exact hardnesses are measured. The Rockwell, Brinell, and Scaleroscope machines give quite definite results which may be easily reproduced at any time. The purpose of this investigation is to determine whether any definite relation exists between Moe's hardness and the hardness as measured by those machines commonly used for the determination of hardness of metals. If such a relation were found it would provide a more definite and accurate measure of the hardness of minerals.

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Contagious bovine pleuropneumonia (CBPP) is a serious respiratory disease of cattle caused by Mycoplasma mycoides subsp. mycoides. Current vaccines against CBPP induce short-lived immunity and can cause severe postvaccine reactions. Previous studies have identified the N terminus of the transmembrane lipoprotein Q (LppQ-N') of M. mycoides subsp. mycoides as the major antigen and a possible virulence factor. We therefore immunized cattle with purified recombinant LppQ-N' formulated in Freund's adjuvant and challenged them with M. mycoides subsp. mycoides. Vaccinated animals showed a strong seroconversion to LppQ, but they exhibited significantly enhanced postchallenge glomerulonephritis compared to the placebo group (P = 0.021). Glomerulonephritis was characterized by features that suggested the development of antigen-antibody immune complexes. Clinical signs and gross pathological scores did not significantly differ between vaccinated and placebo groups. These findings reveal for the first time the pathogenesis of enhanced disease as a result of antibodies against LppQ during challenge and also argue against inclusion of LppQ-N' in a future subunit vaccine for CBPP.

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The voltage-gated sodium channel is the site of action of more than six classes of neurotoxins and drugs that alter its function by interaction with distinct, allosterically coupled receptor sites. Batrachotoxin (BTX) is a steroidal alkaloid that binds to neurotoxin receptor site 2 and causes persistent activation. BTX binding is inhibited allosterically by local anesthetics. We have investigated the interaction of BTX with amino acid residues I1760, F1764, and Y1771, which form part of local anesthetic receptor site in transmembrane segment IVS6 of type IIA sodium channels. Alanine substitution for F1764 (mutant F1764A) reduces tritiated BTX-A-20-α-benzoate binding affinity, causing a 60-fold increase in Kd. Alanine substitution for I1760, which is adjacent to F1764 in the predicted IVS6 transmembrane alpha helix, causes only a 4-fold increase in Kd. In contrast, mutant Y1771A shows no change in BTX binding affinity. For wild-type and mutant Y1771A, BTX shifted the voltage for half-maximal activation ≈40 mV in the hyperpolarizing direction and increased the percentage of noninactivating sodium current to ≈60%. In contrast, these BTX effects were eliminated completely for the F1764A mutant and were reduced substantially for mutant I1760A. Our data suggest that the BTX receptor site shares overlapping but nonidentical molecular determinants with the local anesthetic receptor site in transmembrane segment IVS6 as well as having unique molecular determinants in transmembrane segment IS6, as demonstrated in previous work. Evidently, BTX conforms to a domain–interface allosteric model of ligand binding and action, as previously proposed for calcium agonist and antagonist drugs acting on l-type calcium channels.

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Alternative pre-mRNA splicing patterns can change an extracellular stimulus, but the signaling pathways leading to these changes are still poorly characterized. Here, we describe a tyrosine-phosphorylated nuclear protein, YT521-B, and show that it interacts with the nuclear transcriptosomal component scaffold attachment factor B, and the 68-kDa Src substrate associated during mitosis, Sam68. Northern blot analysis demonstrated ubiquitous expression, but detailed RNA in situ analysis revealed cell type specificity in the brain. YT521-B protein is localized in the nucleoplasm and concentrated in 5–20 large nuclear dots. Deletion analysis demonstrated that the formation of these dots depends on the presence of the amino-terminal glutamic acid-rich domain and the carboxyl-terminal glutamic acid/arginine-rich region. We show that the latter comprises an important protein–protein interaction domain. The Src family kinase p59fyn-mediated tyrosine phosphorylation of Sam68 negatively regulates its association with YT521-B, and overexpression of p59fyn dissolves nuclear dots containing YT521-B. In vivo splicing assays demonstrated that YT521-B modulates alternative splice site selection in a concentration-dependent manner. Together, our data indicate that YT521-B and Sam68 may be part of a signal transduction pathway that influences splice site selection.

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Evernimicin (Evn), an oligosaccharide antibiotic, interacts with the large ribosomal subunit and inhibits bacterial protein synthesis. RNA probing demonstrated that the drug protects a specific set of nucleotides in the loops of hairpins 89 and 91 of 23S rRNA in bacterial and archaeal ribosomes. Spontaneous Evn-resistant mutants of Halobacterium halobium contained mutations in hairpins 89 and 91 of 23S rRNA. In the ribosome tertiary structure, rRNA residues involved in interaction with the drug form a tight cluster that delineates the drug-binding site. Resistance mutations in the bacterial ribosomal protein L16, which is shown to be homologous to archaeal protein L10e, cluster to the same region as the rRNA mutations. The Evn-binding site overlaps with the binding site of initiation factor 2. Evn inhibits activity of initiation factor 2 in vitro, suggesting that the drug interferes with formation of the 70S initiation complex. The site of Evn binding and its mode of action are distinct from other ribosome-targeted antibiotics. This antibiotic target site can potentially be used for the development of new antibacterial drugs.

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Monkeys with excellent reproductive histories were immunized with the laminin peptides YIGSR, RGD, IKVAV, and YD, a control sequence with no known biological function. Sera from the YIGSR-immunized monkey became toxic, causing neural tube defects in whole rat embryo cultures, and this monkey experienced fetal loss after immunization. Sera from the RGD-immunized monkey also became embryotoxic in culture after immunization, but this monkey appeared to become infertile as she failed to initiate a pregnancy for at least 2 years after immunization. In contrast, embryos cultured on sera from the IKVAV- or YD-immunized monkeys were predominantly normal and both monkeys completed successful pregnancies. Antibody levels to the respective peptides or to laminin were not predictive of embryotoxicity, but antibody binding to homogenized yolk sacs as well as to yolk sacs of cultured embryos was associated with sera embryotoxicity and reproductive outcomes in vivo. These observations suggested that the laminin sequences YIGSR and RGD may play a role in immune-mediated reproductive failure by reacting directly with embryonic tissue and could provide a basis for identifying individuals at risk for both spontaneous abortion and infertility.

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The start of accession negotiations between Ankara and the EU is vital for the future of both Turkey and the Union, including Poland as its member state, as well as for the geopolitical situation in Eurasia (the Black Sea region, Caucasus, Central Asia and the Middle East). Appreciating the significance of these issues, the Centre for Eastern Studies in early 2005 decided to launch a project entitled "Turkey after the start of negotiations with the European Union - foreign relations and the domestic situation". The goal of this project is to present, within the context of accession negotiations, Turkey's greatest internal challenges as well as Ankara's relations with its neighbour regions, the EU and the USA. This Report is the first of three which will be published as part of the project. The Report includes texts on Turkish-US relations since 2003, major political and social challenges on Turkey's path towards the EU and the current condition of the Turkish economy. The Report was developed between July 2005 and November 2006, over which time CES workers and associates searched for publicly available materials in Poland, Turkey and EU countries, and went on three research trips to Turkey, where they met local researchers, analysts, politicians and officials. The authors of the Report would like to express their gratitude to everyone who have shared their opinions with them, and to the Polish Embassy in Ankara, especially to Ambassador Grzegorz Michalski and Minister Andrzej Ananicz for their expert support and assistance in the authors' work on this Report. This Report does not present the official stance of the Polish government on the issues discussed therein; instead it reflects the personal views of its authors, who have made their best efforts to ensure that their work is reliable.

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We have investigated if in a cold seep methane or sulfide is used for chemosynthetic primary production and if significant amounts of the sulfide produced by anaerobic oxidation of methane are oxidized geochemically and hence are not available for chemosynthetic production. Geochemically controlled redox reactions and biological turnover were compared in different habitats of the Håkon Mosby Mud Volcano. The center of the mud volcano is characterized by the highest fluid flow, and most primary production by the microbial community depends on oxidation of methane. The small amount of sulfide produced is oxidized geochemically with oxygen or is precipitated with dissolved iron. In the medium flow peripheral Beggiatoa habitat sulfide is largely oxidized biologically. The oxygen and nitrate supply is high enough that Beggiatoa can oxidize the sulfide completely, and chemical sulfide oxidation or precipitation is not important. An internally stored nitrate reservoir with average concentrations of 110 mmol L-1 enables the Beggiatoa to oxidize sulfide anaerobically. The pH profile indicates sequential sulfide oxidation with elemental sulfur as intermediate. Gray thiotrophic mats associated with perturbed sediments showed a high heterogeneity in sulfate turnover and high sulfide fluxes, balanced by the opposing oxygen and nitrate fluxes so that biological oxidation dominates over geochemical sulfide removal processes. The three habitats indicate substantial small-scale variability in carbon fixation pathways either through direct biological use of methane or through indirect carbon fixation of methane-derived carbon dioxide by chemolithotrophic sulfide oxidation.

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Studies of the physical properties of trans-Neptunian objects (TNOs) are a powerful probe into the processes of planetesimal formation and solar system evolution. James Webb Space Telescope (JWST) will provide unique new capabilities for such studies. Here, we outline where the capabilities of JWST open new avenues of investigation, potentially valuable observations and surveys, and conclude with a discussion of community actions that may serve to enhance the eventual science return of JWST's TNO observations.

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Three new bimetallic oxamato-based magnets with the proligand 4,5-dimethyl-1,2-phenylenebis-(oxamato) (dmopba) were synthesized using water or dimethylsulfoxide (DMSO) as solvents. Single crystal X-ray diffraction provided structures for two of them: [MnCu(dmopba)(H(2)O)(3)]n center dot 4nH(2)O (1) and [MnCu(dmopba)(DMSO)(3)](n center dot)nDMSO (2). The crystalline structures for both 1 and 2 consist of linearly ordered oxamato-bridged Mn(II)Cu(II) bimetallic chains. The magnetic characterization revealed a typical behaviour of ferrimagnetic chains for 1 and 2. Least-squares fits of the experimental magnetic data performed in the 300-20 K temperature range led to J(MnCu) = -27.9 cm(-1), g(Cu) = 2.09 and g(Mn) = 1.98 for 1 and J(MnCu) = -30.5 cm(-1), g(Cu) = 2.09 and g(Mn) = 2.02 for 2 (H = -J(MnCu)Sigma S(Mn, i)(S(Cu, i) + S(Cu, i-1))). The two-dimensional ferrimagnetic system [Me(4)N](2n){Co(2)[Cu(dmopba)](3)}center dot 4nDMSO center dot nH(2)O (3) was prepared by reaction of Co(II) ions and an excess of [Cu(dmopba)](2-) in DMSO. The study of the temperature dependence of the magnetic susceptibility as well as the temperature and field dependences of the magnetization revealed a cluster glass-like behaviour for 3.