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61.
62.
Expression of distinct conformations of free HLA-Cw4 heavy chains in transfected neuroblastoma cells
Anna Marozzi Raffaella Meneveri Claudio De Santis Piera Robbioni Elena Molteni Alberto Beretta Antonio G. Siccardi Enrico Ginelli 《Immunogenetics》1996,43(5):289-295
Epitope mapping of HLA-Cw4 indicates that the two monoclonal antibodies (mAbs) L31 and M38, specific for beta 2-microglobulin
(β2m)-free HLA-C heavy chains, react preferentially with the KYK motif, located in the binding groove (α1 domain). Transfection
of HLA-Cw4 cDNA into a neuroblastoma cell line, which normally expresses negligible HLA class I, resulted in the constitutive
surface expression of molecules displaying different reactivities with the two mAbs. This cellular system was used to determine
whether L31 and M38 recognize distinct conformations of β2m-free HLA-C proteins, and to investigate their mechanism of expression. Interferon-γ greatly enhanced the expression of L31-reactive
free chains, while abolishing that of M38-reactive molecules. The cytokine-induced expression of L31-reactive molecules was
inhibited by anti-sense oligonucleotides specific for β2m mRNA, while constitutive expression of L31-reactive molecules was only partially affected. Exogenous β2m resulted in a reduction of constitutive L31 reactivity, and in a concomitant increase of M38 reactivity. These results indicate
that: 1) at the cell surface, L31 and M38 react with two distinct conformations of HLA-Cw4 β2m-free heavy chains, of which the L31-reactive conformation is the least folded; 2) the expression of both conformers can
be modulated by endogenous or exogenous β2m; and (3) L31-reactive molecules exposed at the cell surface are likely to derive from the dissociation of empty HLA-Cw4/β2m complexes. 相似文献
63.
Phosphorylation of the Casein Kinase II Domain of B-50 (GAP-43) in Rat Cortical Growth Cones 总被引:1,自引:1,他引:0
†Michael A. N. Edgar ‡Piera Pasinelli M. DeWit §Brian Anton †Linda A. Dokas Lucia Pastorino Monica DiLuca Flaminio Cattabeni Willem H. Gispen Pierre N. E. De Graan 《Journal of neurochemistry》1997,69(5):2206-2215
Abstract: Growth-associated phosphoprotein B-50 is a neural protein kinase C (PKC) substrate enriched in nerve growth cones that has been implicated in growth cone plasticity. Here we investigated whether B-50 is a physiological substrate for casein kinase II (CKII) in purified rat cortical growth cone preparations. Using site-specific proteolysis and known modulators of PKC, in combination with immunoprecipitation, mass spectrometry, and phosphoamino acid analysis, we demonstrate that endogenous growth cone B-50 is phosphorylated at multiple sites, on both serine and threonine residues. Consistent with previous reports, stimulation of PKC activity increased the phosphorylation of only those proteolytic fragments containing Ser41 . Under basal conditions, however, phosphorylation was predominantly associated with fragments not containing Ser41 . Mass spectrometry of tryptic digests of B-50, which had been immunoprecipitated from untreated growth cones, revealed that in situ phosphorylation occurs within peptides B-50181–198 and B-5082–98 . These peptides contain the major and minor in vitro CKII phosphosites, respectively. In addition, cyanogen bromide digestion of immunoprecipitated chick B-50 generated a 4-kDa C-terminal B-50 phosphopeptide, confirming that phosphorylation of the CKII domain occurs across evolutionary diverse species. We conclude that B-50 in growth cones is not only a substrate for PKC, but also for CKII. 相似文献
64.
Luca Bragazza Juul Limpens† Renato Gerdol Philippe Grosvernier‡ Michal Hájek§ Tomá Hájek¶ Petra Hajkova§ Ina Hansen Paola Iacumin Lado Kutnar†† Håkan Rydin‡‡ Teemu Tahvanainen§§ 《Global Change Biology》2005,11(1):106-114
Alteration of the global nitrogen (N) cycle because of human‐enhanced N fixation is a major concern particularly for those ecosystems that are nutrient poor by nature. Because Sphagnum‐dominated mires are exclusively fed by wet and dry atmospheric deposition, they are assumed to be very sensitive to increased atmospheric N input. We assessed the consequences of increased atmospheric N deposition on total N concentration, N retention ability, and δ15N isotopic signature of Sphagnum plants collected in 16 ombrotrophic mires across 11 European countries. The mires spanned a gradient of atmospheric N deposition from about 0.1 up to about 2 g m?2 yr?1. Mean N concentration in Sphagnum capitula was about 6 mg g?1 in less polluted mires and about 13 mg g?1 in highly N‐polluted mires. The relative difference in N concentration between capitulum and stem decreased with increasing atmospheric N deposition, suggesting a possible metabolic mechanism that reduces excessive N accumulation in the capitulum. Sphagnum plants showed lower rates of N absorption under increasing atmospheric N deposition, indicating N saturation in Sphagnum tissues. The latter probably is related to a shift from N‐limited conditions to limitation by other nutrients. The capacity of the Sphagnum layer to filter atmospheric N deposition decreased exponentially along the depositional gradient resulting in enrichment of the mire pore water with inorganic N forms (i.e., NO3?+NH4+). Sphagnum plants had δ15N signatures ranging from about ?8‰ to about ?3‰. The isotopic signatures were rather related to the ratio of reduced to oxidized N forms in atmospheric deposition than to total amount of atmospheric N deposition, indicating that δ15N signature of Sphagnum plants can be used as an integrated measure of δ15N signature of atmospheric precipitation. Indeed, mires located in areas characterized by greater emissions of NH3 (i.e., mainly affected by agricultural activities) had Sphagnum plants with a lower δ15N signature compared with mires located in areas dominated by NOx emissions (i.e., mainly affected by industrial activities). 相似文献
65.
Microsomal vesicles from 24-hour-old radish (Raphanus sativus L.) seedlings accumulate Ca2+ upon addition of MgATP. MgATP-dependent Ca2+ uptake co-migrates with the plasma membrane H+-ATPase on a sucrose gradient. Ca2+ uptake is insensitive to oligomycin, inhibited by vanadate (IC50 40 micromolar) and erythrosin B (IC50 0.2 micromolar) and displays a pH optimum between pH 6.6 and 6.9. MgATP-dependent Ca2+ uptake is insensitive to protonophores. These results indicate that Ca2+ transport in these microsomal vesicles is catalyzed by a Mg2+-dependent ATPase localized on the plasma membrane. Ca2+ strongly reduces ΔpH generation by the plasma membrane H+-ATPase and increases MgATP-dependent membrane potential difference (Δψ) generation. These effects of Ca2+ on ΔpH and Δψ generation are drastically reduced by micromolar erythrosin B, indicating that they are primarily a consequence of Ca2+ uptake into plasma membrane vesicles. The Ca2+-induced increase of Δψ is collapsed by permeant anions, which do not affect Ca2+-induced decrease of ΔpH generation by the plasma membrane H+-ATPase. The rate of decay of MgATP-dependent ΔpH, upon inhibition of the plasma membrane H+-ATPase, is accelerated by MgATP-dependent Ca2+ uptake, indicating that the decrease of ΔpH generation induced by Ca2+ reflects the efflux of H+ coupled to Ca2+ uptake into plasma membrane vesicles. It is therefore proposed that Ca2+ transport at the plasma membrane is mediated by a Mg2+-dependent ATPase which catalyzes a nH+/Ca2+ exchange. 相似文献
66.
Giuseppina Mazzocchi Ludwig K. Malendowicz Piera Rebuffat Claudia Robba Giuseppe Gottardo Prof. Gastone G. Nussdorfer 《Cell and tissue research》1986,243(2):303-310
Summary Short-term ACTH treatment provoked a decrease in volume of the lipid-droplet compartment in rat zona glomerulosa cells, and a rise in plasma and intracellular concentrations of corticosterone and aldosterone. It enhanced activities of 3-hydroxysteroid dehydrogenase (3HSD), 11-hydroxylase (11OH) and 18-hydroxylase (18OH). Long-term ACTH administration produced a hypertrophy of the zona glomerulosa and its parenchymal cells, a result of the increase in volume of the smooth endoplasmic reticulum and the mitochondrial compartment. The surface area per cell of mitochondrial inner membranes increased; the tubular cristae were transformed into a homogeneous population of vesicles. The plasma and intracellular concentrations of corticosterone further increased, whereas those of aldosterone fell below basal levels (the aldosterone-escape phenomenon). The activities of 3HSD and 11OH were enhanced, that of 180H decreased. Therefore, ACTH stimulates zona glomerulosa growth and transforms parenchymal elements into zona fasciculata celltypes. Cyanoketone nullified acute ACTH effects on plasma and intracellular concentrations of corticosterone and aldosterone, but did not affect the activities of 11OH and 18OH. Chronic ACTH treatment produced similar results, although 18OH activity was not suppressed. The mechanism underlying the aldosterone-escape phenomenon may thus involve a rise in the intracellular concentration of corticosterone, caused by the enhanced synthesis and activation of 3HSD and 11OH. 相似文献
67.
The iron status of 50 patients with Ph'-positive chronic granulocytic leukaemia (CGL) was evaluated at diagnosis by means of bone marrow and blood studies. A decreased or absent iron in semiquantitative estimation on bone marrow smears was observed in 92% of patients, and 88% had a low sideroblast score. In contrast, normal Hb and serum iron concentrations were found in the majority of cases, and only two out of the 50 patients displayed a decreased serum ferritin. To ascertain whether the bone marrow pattern of iron depletion could be due to an expansion of the red cell mass, the latter parameter was measured by isotopic methods in a subgroup of 11 patients. Normal or slightly increased values were obtained in all cases. We conclude that absent or decreased marrow iron is a common feature in the chronic phase of CGL, that generally does not reflect true iron deficiency. Since such a finding is also usual in polycythaemia vera and idiopathic myelofibrosis, it should be included among the features shared by the chronic myeloproliferative disorders. 相似文献
68.
69.
The iron status of 26 patients with essential thrombocythaemia (ET) was evaluated at diagnosis by means of bone marrow iron and blood studies, including serum ferritin determination. Nine patients were males, 17 females, and the mean age was 53 years (range 7-81). A decreased or absent iron level by semiquantitative estimation on bone marrow smears was observed in 77% of patients, and 81% had a low sideroblast score. Such a marrow pattern of iron depletion was equally distributed between both sexes. Contrasting with this, normal Hb, MCV, serum iron and serum ferritin were registered in the majority of cases. According to these results, absent or decreased marrow iron would be a common feature in ET, generally not reflecting true iron deficiency, as it occurs in the remaining chronic myeloproliferative disorders. Thus, in patients in whom ET is suspected, the diagnostic criterion of ruling out iron deficiency would be better served by serum ferritin measurement than by bone marrow iron estimation. 相似文献
70.
Pineal cell aggregates in 5, 10 and 15 day-old chick embryos have been studied. Cell aggregates were classified into rosettes or vesicles and spheroid and ellipsoid vesicles distinguished. The number of pineal vesicles per unit of surface (vesicle density) was determined in three pineal portions: apical, anterior and posterior. By day 5, only cellular rosettes were found, mainly in the apical portion. After 10 and 15 days, the presence of rosettes was occasional. The posterior wall showed only small spheroid vesicles, while in the apical and anterior areas ellipsoid vesicles were also observed. Moreover, the spheroid/ellipsoid vesicle ratio increased from the 10th to the 15th day of incubation. The vesicle density decreased between the 10th and 15th day because of the increase in both vesicle and pineal size, without changes in the total number of vesicles. The results suggest that changes in vesicle morphology and density could be related to the functional activity of the pineal gland during embryonic development. 相似文献