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Internal fatty acylation of proteins is a recognized means of modifying biological behavior. Escherichia coli hemolysin A (HlyA), a toxic protein, is transcribed as a nontoxic protein and made toxic by internal acylation of two lysine residue epsilon-amino groups; HlyC catalyzes the acyl transfer from acyl-acyl carrier protein (ACP), the obligate acyl donor. Conserved residues among the respective homologous C proteins that activate 13 different RTX (repeats in toxin) toxins of which HlyA is the prototype likely include some residues that are important in catalysis. Possible roles of two conserved tyrosines and two conserved arginines were investigated by noting the effects of chemical modifiers and site-directed mutagenesis. TNM modification of HlyC at pH 8.0 led to extensive inhibition that was prevented by the presence of the substrate myristoyl-ACP but not by the product, ACPSH. NAI had no effect. Y70G and Y150G greatly diminished enzyme activity, whereas mutations Y70F and Y150F exhibited wild-type activity. Modification of arginine residues with PG markedly lowered acyltransferase activity with moderate protection by both myristoyl-ACP and ACPSH. Under optimum conditions, four separate mutations of the two conserved arginine residues (R24A, R24K, R87A, and R87K) had little effect on acyltransferase activity. 相似文献
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The Annual bioProcessUK Conference has acted as the key networking event for bioprocess scientists and engineers in the UK for the past 10 years. The following article is a report from the sessions that focused on continuous bioprocessing during the 10th Annual bioProcessUK Conference (London, December 2013). These sessions were organized by the ‘EPSRC Centre for Innovative Manufacturing in Emergent Macromolecular Therapies’ hosted at University College London. A plenary lecture and workshop provided a forum for participants to debate topical issues in roundtable discussions with industry and academic experts from institutions such as Genzyme, Janssen, Novo Nordisk, Pfizer, Merck, GE Healthcare and University College London. The aim of these particular sessions was to understand better the challenges and opportunities for continuous bioprocessing in the bioprocessing sector. 相似文献
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M D Keppler M A Read P J Perry J O Trent T C Jenkins A P Reszka S Neidle K R Fox 《European journal of biochemistry》1999,263(3):817-825
We have used quantitative DNase I footprinting to measure the relative affinities of four disubstituted and two monosubstituted amidoanthraquinone compounds for intermolecular DNA triplexes, and have examined how the position of the attached base-functionalized substituents affects their ability to stabilize DNA triplexes. All four isomeric disubstituted derivatives examined stabilize DNA triplexes at micromolar or lower concentrations. Of the compounds studied the 2,7-disubstituted amidoanthraquinone displayed the greatest triplex affinity. The order of triplex affinity for the other disubstituted ligands decreases in the order 2,7 > 1,8 = 1,5 > 2,6, with the equivalent monosubstituted compounds being at least an order of magnitude less efficient. The 1,5-disubstituted derivative also shows some interaction with duplex DNA. These results have been confirmed by molecular modelling studies, which provide a rational basis for the structure-activity relationships. These suggest that, although all of the compounds bind through an intercalative mode, the 2,6, 2,7 and 1,5 disubstituted isomers bind with their two side groups occupying adjacent triplex grooves, in contrast with the 1,8 isomer which is positioned with both side groups in the same triplex groove. 相似文献
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At a high arctic lowland on Ellesmere Island, heath communities dominated by Cassiope tetragona were widespread, and occurred in a variety of habitats that differred in time of snowmelt, relative site moisture, soil thaw depth, and air and soil temperature. Cover, standing crop, and production were dominated by woody plants, notably Cassiope tetragona and Dryas integrifolia . The Cassiope -dominated heaths were similar in composition to those at other Canadian high arctic localities, but were less similar to localities in Greenland and Spitzbergen. Compositional relationships among Cassiope -dominated heaths in different habitats at the study site may be largely determined by two interrelated environmental factors, time of snowmelt and site moisture.
Aboveground vascular plant biomass was concentrated near the ground surface, resulting in simple vertical structure that takes advantage of relatively warm soil and air temperatures in summer, and a protective snow cover in winter. Aboveground vascular plant standing crop was largely comprised of attached dead tissue, and there were equal proportions of above- and belowground biomass. Low community production was due to dominance by long-lived, slow-growing species, and to short growing seasons and cold temperatures. Overall character of Cassiope -dominated heaths reflects the conservative, stress tolerant growth strategy of the dominant species, Cassiope tetragona . 相似文献
Aboveground vascular plant biomass was concentrated near the ground surface, resulting in simple vertical structure that takes advantage of relatively warm soil and air temperatures in summer, and a protective snow cover in winter. Aboveground vascular plant standing crop was largely comprised of attached dead tissue, and there were equal proportions of above- and belowground biomass. Low community production was due to dominance by long-lived, slow-growing species, and to short growing seasons and cold temperatures. Overall character of Cassiope -dominated heaths reflects the conservative, stress tolerant growth strategy of the dominant species, Cassiope tetragona . 相似文献
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Summary Some types of nondeletional heterocellular hereditary persistence of fetal hemoglobin (HPFH) appear to be caused by mutations in the globin gene cluster near the globin genes, while in other cases the condition is associated with a gene or genes outside the globin gene complex. We have used DNA probes for chromosome 11 markers to localize the HPFH determinant in a large Australian kindred with nondeletional heterocellular HPFH. In 13 of the 58 family members studied the Hb F levels are increased to between 1.8% and 7.9%, the Hb F being composed predominantly of A chains. All family members were typed for restriction fragment length polymorphisms detected by probes from the globin gene complex, and the nearby genetic markers D11S12, INS, and HRAS. Linkage analysis showed HPFH is closely linked to the globin gene cluster (confidence limits of 0,0.0-0.19), D11S12 (0, 0.0-0.23) and the insulin gene (0,0.0-0.11). These data and the chain composition are consistent with HPFH in this family being caused by a mutation within the globin gene cluster. 相似文献