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81.
M?ssbauer spectra from frozen solutions of ovotransferrin were recorded in a variety of applied external magnetic fields and at various temperatures in a small applied field. The results were fitted to a simple model for the electronic structure at the iron site. This model requires admixtures of the free ion 6S and 4P states, indicating a weak cubic crystal field. Possible implications of this model regarding the binding site are discussed. 相似文献
82.
The binding of Ga(III) to the proteolytically derived N-terminal half-molecule of human transferrin (HTF/2N) was studied by proton nuclear magnetic resonance spectroscopy. The pH-dependent titration curves of the histidinyl C(2) proton chemical shifts were altered upon formation of the GaIIIHTF/2N(C2O4) ternary complex. Two high-pK'a histidines failed to titrate when the metal and synergistic anion formed a complex with the protein. These results implicated two histidinyl residues as direct ligands to the metal. The rates of hydrogen-deuterium exchange for the C(2) protons of certain histidinyl residues were substantially decreased by metal ion binding. The two ligand histidines were protected from exchange, and a third, low-pK'a, histidinyl residue was protected. We propose that this third histidinyl residue is involved in anion binding and may serve as the base in the putative proton-relay scheme proposed for complex formation. 相似文献
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Bradley K. Woodworth Charles M. Francis Philip D. Taylor 《Journal of avian biology》2014,45(4):387-395
Inland dispersal of migrating land birds away from the coast, often opposite to the direction of migration, occurs frequently. Many of these movements may involve migrants seeking improved stopover conditions farther inland, but direct study of inland flights and of the ecological factors influencing their occurrence is limited. We used an automated telemetry array and ground‐tracking to assess flight behaviours, survival, and habitat use of young red‐eyed vireos Vireo olivaceus during fall migration at a coastal island and an inland stopover site in southwest Nova Scotia. We recorded inland flights for 41% (11/27) of individuals that departed the island. At least 25% of 16 individuals tagged at the inland site also relocated within the landscape prior to continuing migration, but due to the higher proportion of ambiguous flights at the inland site (44%) compared to the island (15%), we could not be sure if actual proportions of relocations differed between sites. Mortality on the island (at least 10 of 39 individuals) was significantly higher than at the inland site (0 of 16 individuals). At mainland sites near the coast where we found 6 of 11 individuals after they relocated away from the island, mortality remained high (2/6). Lack of deciduous canopy cover may have contributed to the high mortality on the island, but coastal mainland sites had a relatively high amount of deciduous canopy cover, similar to at the inland site where there was no mortality. Although coastal stopover sites may be important for migrating songbirds, especially before or after making a large water crossing, our results show that mortality can be much higher, and habitat poorer, at the coast compared to farther inland. Therefore, relocating inland may be an adaptive strategy for individuals that initially settle at the coast and that need to rest and refuel before they continue migration. 相似文献
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Alejandra Garces Krishnamohan Atmakuri Michael R. Chase Joshua S. Woodworth Bryan Krastins Alissa C. Rothchild Talia L. Ramsdell Mary F. Lopez Samuel M. Behar David A. Sarracino Sarah M. Fortune 《PLoS pathogens》2010,6(6)
Mycobacterium tuberculosis (Mtb) requires the ESX1 specialized protein secretion system for virulence, for triggering cytosolic immune surveillance pathways, and for priming an optimal CD8+ T cell response. This suggests that ESX1 might act primarily by destabilizing the phagosomal membrane that surrounds the bacterium. However, identifying the primary function of the ESX1 system has been difficult because deletion of any substrate inhibits the secretion of all known substrates, thereby abolishing all ESX1 activity. Here we demonstrate that the ESX1 substrate EspA forms a disulfide bonded homodimer after secretion. By disrupting EspA disulfide bond formation, we have dissociated virulence from other known ESX1-mediated activities. Inhibition of EspA disulfide bond formation does not inhibit ESX1 secretion, ESX1-dependent stimulation of the cytosolic pattern receptors in the infected macrophage or the ability of Mtb to prime an adaptive immune response to ESX1 substrates. However, blocking EspA disulfide bond formation severely attenuates the ability of Mtb to survive and cause disease in mice. Strikingly, we show that inhibition of EspA disulfide bond formation also significantly compromises the stability of the mycobacterial cell wall, as does deletion of the ESX1 locus or individual components of the ESX1 system. Thus, we demonstrate that EspA is a major determinant of ESX1-mediated virulence independent of its function in ESX1 secretion. We propose that ESX1 and EspA play central roles in the virulence of Mtb in vivo because they alter the integrity of the mycobacterial cell wall. 相似文献
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Comparative studies of the binding and growth-supportive ability of mammalian transferrins in human cells 总被引:1,自引:0,他引:1
The ability of human-derived cells in culture to bind, remove iron from, and grow in the presence of transferrins (Tf) isolated from the sera of species commonly included in tissue culture medium was investigated. Kinetic studies on HeLa cells reveal apparent first-order association rate constants of 0.43 min-1 for human Tf and 0.15 min-1 for equine Tf. Labeled chicken ovo-Tf and fetal bovine Tf were not recognized by the HeLa cells. Competition experiments with HeLa cells that use either isolated Tf or parent serum confirm these findings. Equilibrium binding experiments performed on HeLa cells at 37 degrees C in the presence of 2,4-dinitrophenol to prevent iron removal indicate 1 X 10(6) Tf bound/cell with a dissociation constant (K'D) of 28 nM for human Tf and 182 nM for equine Tf. Equilibrium binding performed at 0 degrees C to prevent endocytosis reveals 4.1-6.7 X 10(5) Tf binding sites/cell with a K'D of 8.3 nM for human Tf and 41.5 nM for equine Tf. Parallel experiments in normal human diploid fibroblast-like MRC-5 cells indicate expression of 0.82-2.78 X 10(5) Tf binding sites/cell with a K'D of 8.2 nM for human and 39.1 nM for equine Tf. Thus, the results of equilibrium binding studies of a more differentiated cell type are consistent with those found for HeLa cells. Fetal bovine Tf was found to compete weakly with labeled human Tf for human receptor on HeLa cells in a soluble receptor assay, with an approximately 500-fold excess needed to reduce binding to half maximal. Iron uptake experiments show an iron donating hierarchy where human greater than horse greater than calf, suggesting that the rate of iron uptake depends on the affinity of receptor for transferrin. Growth experiments involving HeLa cells in chemically defined serum-free medium demonstrate that bovine Tf will support growth as well as human Tf, but at concentrations much higher than are required of human Tf. 相似文献
90.
Characterization of the Yucatan miniature pig skin and small intestine for pharmaceutical applications 总被引:4,自引:0,他引:4
Yucatan miniature pig skin and gastrointestinal tract have been characterized as potential models for human organ systems in in vitro and in vivo pharmaceutical experiments. Histology of pig skin and the flux of one classical drug, caffeine, are described. The Yucatan small intestine was examined morphologically. Both Yucatan skin and GI tract were found to have similarities to man. 相似文献