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排序方式: 共有883条查询结果,搜索用时 15 毫秒
1.
Subunit composition and ATP site labeling of the coated vesicle proton-translocating adenosinetriphosphatase 总被引:3,自引:0,他引:3
The partially purified proton-translocating adenosinetriphosphatase [(H+)-ATPase] from clathrin-coated vesicles has been reported to contain eight polypeptides of molecular weights 15,000-116,000 [Xie, X.S., & Stone, D.K. (1986) J. Biol. Chem. 261, 2492-2495]. To determine whether these polypeptides form a single macromolecular complex, we have isolated three monoclonal antibodies which recognize the reconstitutively active (H+)-ATPase in the native, detergent-solubilized state. All three monoclonal antibodies precipitate the same set of polypeptides from either the partially purified enzyme or the detergent-solubilized coated vesicle membrane proteins. The immunoprecipitated polypeptides have molecular weights of 100,000, 73,000, 58,000, 40,000, 38,000, 34,000, 33,000, 19,000, and 17,000. These results thus indicate that this set of polypeptides forms a single macromolecular complex and suggest that they correspond to subunits of the coated vesicle (H+)-ATPase. To identify the ATP-hydrolytic subunit of the coated vesicle (H+)-ATPase, the purified enzyme was reacted with N-ethylmaleimide (NEM) and 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl), both of which inhibit activity in an ATP-protectable manner. Labeling was carried out by using [3H]NEM or [14C]NBD-Cl, and the specificity of the reaction was increased by prelabeling of the protein with the nonradioactive reagents in the presence of ATP and by taking advantage of the nucleotide specificity of protection. The principal polypeptide labeled by both [3H]NEM and [14C]NBD-Cl had a molecular weight of 73,000. In addition, this protein was the only polypeptide whose labeling was significantly reduced in the presence of ATP.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
2.
Adriana Calderaro Giovanna Piccolo Chiara Gorrini Sara Montecchini Sabina Rossi Maria Cristina Medici Carlo Chezzi Georges Snounou 《PloS one》2012,7(10)
It has been proposed that ovale malaria in humans is caused by two closely related but distinct species of malaria parasites: P. ovale curtisi and P. ovale wallikeri. We have extended and optimized a Real-time PCR assay targeting the parasite’s small subunit ribosomal RNA (ssrRNA) gene to detect both these species. When the assay was applied to 31 archival blood samples from patients diagnosed with P. ovale, it was found that the infection in 20 was due to P. ovale curtisi and in the remaining 11 to P. ovale wallikeri. Thus, this assay provides a useful tool that can be applied to epidemiological investigations of the two newly recognized distinct P. ovale species, that might reveal if these species also differ in their clinical manifestation, drugs susceptibility and relapse periodicity. The results presented confirm that P. ovale wallikeri is not confined to Southeast Asia, since the majority of the patients analyzed in this study had acquired their P. ovale infection in African countries, mostly situated in West Africa. 相似文献
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Resistance of primary CD8+ cytotoxic T lymphocytes to lysis by cytotoxic granules from cloned T cell lines 总被引:3,自引:0,他引:3
C Nagler-Anderson C R Verret A A Firmenich M Berne H N Eisen 《Journal of immunology (Baltimore, Md. : 1950)》1988,141(10):3299-3305
Recent evidence has shown that cloned, murine CTL cell lines are resistant to the cytotoxic components of the toxic granules they release upon specific interaction with their target cells. Inasmuch as the resistance might be due to selection in culture over many months by repeated exposure to these cytolytic components (which are released repeatedly as a result of the cultured CTL being periodically stimulated by target cells), we asked whether primary CTL are also resistant. The primary CTL were elicited in vivo by i.p. injection of allogeneic tumor cells or in vitro by 5- to 6-day MLC or by 48-h exposure to the lectin Con A. The responding cells were separated into purified CD8+ (i.e., CD4-, CD8+) and purified CD4+ (i.e., CD4+, CD8-) T cell populations that were analyzed for cytolytic activity and for resistance to lysis by toxic secretory granules derived from cloned CTL cell lines. The CD8+ T cells were highly cytolytic and relatively resistant; they retained their cytolytic activity and were lysed to a minimal extent (0 to 10%) by quantities of isolated granules that lysed 80 to 90% of the P815 tumor cell line (tested as a representative standard cell line). The CD4+ T cells, in contrast, had only minimal cytolytic activity and were far more susceptible to granule-mediated lysis. Although the resistance of primary CD8+ T cells is impressive, it is not as pronounced as the resistance of the cloned CTL cell lines, indicating that during long-term culture there is some selection for increased resistance to granule-mediated lysis. In contrast to T cells (especially CD8+ T cells), Ia+ macrophages, isolated from primary immune peritoneal exudates, were highly susceptible to granule-mediated lysis. 相似文献
6.
Elisabeth Oppliger Leibundgut Bendicht Wermuth Jean-Pierre Colombo Sabina Liechti-Gallati 《Human genetics》1996,97(2):209-213
Ornithine transcarbamylase (OTC) deficiency, the most common inborn error of the urea cycle, shows X-linked inheritance with frequent new mutations. Using polymerase chain reaction (PCR) amplification of the individual exons including adjacent intron sequences followed by direct sequencing of the amplimers we identified four new mutations affecting donor splice sites of introns 2, 5, 6, and 8. The mutation at the first position of intron 2 was a G to A exchange associated with acute neonatal hyperammonemia in a male patient at the age of 5 months. A G to C substitution in intron 5 was detected in a boy who developed 2 days after birth hypotonia, and respiratory distress, followed by severe hyperammonemia and terminal coma. The intron 6 mutation, a G to T substitution, was detected in a girl presenting with first episodes of vomiting and agitation at the age of 2 months. The mutation in intron 8, also a G to T transition, caused fatal hyperammonemia and early death at the age of 15 days in a male patient. We present four donor splice site mutations resulting in severe neonatal or very early onset of the disease in three boys and in one female patient. As the GT dinucleotide of the 5 donor splice site is invariant and required for correct splicing the described mutations may lead to improperly spliced mRNAs and aberrant gene products. 相似文献
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8.
Modifications in the exposure to the solvent of hydrophobic residues, changes in their organization into surface hydrophobic patches, and alterations in the dimerization equilibrium ofβ-lactoglobulin upon thermal treatment at neutralpH were studied. Exposure of tryptophan residues was temperature dependent and was essentially completed on the time scale of seconds. Reorganization of generic hydrophobic protein patches on the protein surface was monitored through binding of 1,8-anilinonaphthalenesulfonate, and was much slower than changes in tryptophan exposure. Different phases in surface hydrophobicity changes were related to the swelling and the subsequent collapse of the protein, which formed a metastable swollen intermediate. Heat treatment ofβ-lactoglobulin also resulted in the formation of soluble oligomeric aggregates. The aggregation process was studied as a function of temperature, demonstrating that (i) dimer dissociation was a necessary step in a sequential polymerization mechanism and (ii) cohesion of hydrophobic patches was the major driving force for aggregation. 相似文献
9.
Graham J. Hymus David P. Johnson Sabina Dore† Hans P. Anderson C. Ross Hinkle‡ Bert G. Drake 《Global Change Biology》2003,9(12):1802-1812
We report the results of a 2‐year study of effects of the elevated (current ambient plus 350 μmol CO2 mol?1) atmospheric CO2 concentration (Ca) on net ecosystem CO2 exchange (NEE) of a scrub–oak ecosystem. The measurements were made in open‐top chambers (OTCs) modified to function as open gas‐exchange systems. The OTCs enclosed samples of the ecosystem (ca. 10 m2 surface area) that had regenerated after a fire, 5 years before, in either current ambient or elevated Ca. Throughout the study, elevated Ca increased maximum NEE (NEEmax) and the apparent quantum yield of the NEE (φNEE) during the photoperiod. The magnitude of the stimulation of NEEmax, expressed per unit ground area, was seasonal, rising from 50% in the winter to 180% in the summer. The key to this stimulation was effects of elevated Ca, and their interaction with the seasonal changes in the environment, on ecosystem leaf area index, photosynthesis and respiration. The separation of these factors was difficult. When expressed per unit leaf area the stimulation of the NEEmax ranged from 7% to 60%, with the increase being dependent on increasing soil water content (Wsoil). At night, the CO2 effluxes from the ecosystem (NEEnight) were on an average 39% higher in elevated Ca. However, the increase varied between 6% and 64%, and had no clear seasonality. The partitioning of NEEnight into its belowground (Rbelow) and aboveground (Rabove) components was carried out in the winter only. A 35% and 27% stimulation of NEEnight in December 1999 and 2000, respectively, was largely due to a 26% and 28% stimulation of Rbelow in the respective periods, because Rbelow constituted ca. 87% of NEEnight. The 37% and 42% stimulation of Rabove in December 1999 and 2000, respectively, was less than the 65% and 80% stimulation of the aboveground biomass by elevated Ca at these times. An increase in the relative amount of the aboveground biomass in woody tissue, combined with a decrease in the specific rate of stem respiration of the dominant species Quercus myrtifolia in elevated Ca, was responsible for this effect. Throughout this study, elevated Ca had a greater effect on carbon uptake than on carbon loss, in terms of both the absolute flux and relative stimulation. Consequently, for this scrub–oak ecosystem carbon sequestration was greater in the elevated Ca during this 2‐year study period. 相似文献
10.
Michael Lippert Karl Steiner Hans-Dieter Payer Sabina Simons Christian Langebartels Heinrich Sandermann Jr. 《Trees - Structure and Function》1996,10(4):268-275
An exposure — response study with proportionalto-ambient ozone levels was conducted in closed chambers on 3-year-old European beech (Fagus sylvatica L.) of montane origin. The fumigation started in April 1990 and lasted for a single growing season. Climate data and ozone concentrations monitored at an experimental station of the Institute for Applied Plant Biology, Schönenbuch, Switzerland were simulated in the exposure chambers 12 days later (1*O3). To test exposure-response relations three additional treatments were applied, subambient (0.2*O3) and two proportionally increased ozone treatments (1.5*O3 and 2*O3). The photosynthetic behaviour of the trees in August revealed the light reactions to be less affected than parameters which are related to the dark reactions of photosynthesis. Assimilation (A350), apparent carboxylation efficiency (CE), and maximum photosynthetic capacity (A2500) were reduced with increasing ozone concentration. For the ozone response of CE and A2500 Critical Levels were calculated. 相似文献