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排序方式: 共有123条查询结果,搜索用时 31 毫秒
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Chanroj S Lu Y Padmanaban S Nanatani K Uozumi N Rao R Sze H 《The Journal of biological chemistry》2011,286(39):33931-33941
The complexity of intracellular compartments in eukaryotic cells evolved to provide distinct environments to regulate processes necessary for cell proliferation and survival. A large family of predicted cation/proton exchangers (CHX), represented by 28 genes in Arabidopsis thaliana, are associated with diverse endomembrane compartments and tissues in plants, although their roles are poorly understood. We expressed a phylogenetically related cluster of CHX genes, encoded by CHX15-CHX20, in yeast and bacterial cells engineered to lack multiple cation-handling mechanisms. Of these, CHX16-CHX20 were implicated in pH homeostasis because their expression rescued the alkaline pH-sensitive growth phenotype of the host yeast strain. A smaller subset, CHX17-CHX19, also conferred tolerance to hygromycin B. Further differences were observed in K(+)- and low pH-dependent growth phenotypes. Although CHX17 did not alter cytoplasmic or vacuolar pH in yeast, CHX20 elicited acidification and alkalization of the cytosol and vacuole, respectively. Using heterologous expression in Escherichia coli strains lacking K(+) uptake systems, we provide evidence for K(+) ((86)Rb) transport mediated by CHX17 and CHX20. Finally, we show that CHX17 and CHX20 affected protein sorting as measured by carboxypeptidase Y secretion in yeast mutants grown at alkaline pH. In plant cells, CHX20-RFP co-localized with an endoplasmic reticulum marker, whereas RFP-tagged CHX17-CHX19 co-localized with prevacuolar compartment and endosome markers. Together, these results suggest that in response to environmental cues, multiple CHX transporters differentially modulate K(+) and pH homeostasis of distinct intracellular compartments, which alter membrane trafficking events likely to be critical for adaptation and survival. 相似文献
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Plants pass the salt 总被引:16,自引:0,他引:16
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Aranda-Sicilia MN Cagnac O Chanroj S Sze H Rodríguez-Rosales MP Venema K 《Biochimica et biophysica acta》2012,1818(9):2362-2371
KEA genes encode putative K(+) efflux antiporters that are predominantly found in algae and plants but are rare in metazoa; however, nothing is known about their functions in eukaryotic cells. Plant KEA proteins show homology to bacterial K(+) efflux (Kef) transporters, though two members in the Arabidopsis thaliana family, AtKEA1 and AtKEA2, have acquired an extra hydrophilic domain of over 500 residues at the amino terminus. We show that AtKEA2 is highly expressed in leaves, stems and flowers, but not in roots, and that an N-terminal peptide of the protein is targeted to chloroplasts in Arabidopsis cotyledons. The full-length AtKEA2 protein was inactive when expressed in yeast; however, a truncated AtKEA2 protein (AtsKEA2) lacking the N-terminal domain complemented disruption of the Na(+)(K(+))/H(+) antiporter Nhx1p to confer hygromycin resistance and tolerance to Na(+) or K(+) stress. To test transport activity, purified truncated AtKEA2 was reconstituted in proteoliposomes containing the fluorescent probe pyranine. Monovalent cations reduced an imposed pH gradient (acid inside) indicating AtsKEA2 mediated cation/H(+) exchange with preference for K(+)=Cs(+)>Li(+)>Na(+). When a conserved Asp(721) in transmembrane helix 6 that aligns to the cation binding Asp(164) of Escherichia coli NhaA was replaced with Ala, AtsKEA2 was completely inactivated. Mutation of a Glu(835) between transmembrane helix 8 and 9 in AtsKEA2 also resulted in loss of activity suggesting this region has a regulatory role. Thus, AtKEA2 represents the founding member of a novel group of eukaryote K(+)/H(+) antiporters that modulate monovalent cation and pH homeostasis in plant chloroplasts or plastids. 相似文献
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Tripković B Buković D Sakić K Sakić S Buković N Radaković B 《Collegium antropologicum》2008,32(1):153-160
Several methods have been found to be successful in reducing the need for allogeneic transfusion among the patients undergoing total hip replacement. The purpose of this prospective study was to analyse the quality and evaluate the effect of postoperative autotransfusion on the need for allogeneic transfusion following total hip replacement. The prospective study was performed in two groups of patients undergoing total hip replacement. Before the operative procedure all patients in both groups predonated two doses of autologous blood. In GROUP 1. the system for postoperative collection and transfusion of shed blood was used. In GROUP 2. the patients underwent total hip replacement without blood salvage system. Standard suction collection sets were used postoperatively. In this group shed blood was not transfused to the patients. The samples of preoperative donated autologus blood, allogeneic blood and postoperative collected autologous blood were analysed for number of red cells, hemoglobin, hematocrit, platelets, white blood cells, values of potassium, sodium, free hemoglobin and acid base status. The postoperatively blood salvage significantly reduced the use of allogeneic transfusion among patients managed with total hip replacement (allogeneic transfusion received 12% patients in Group 1 and 80% patients in Group 2; p<0.001). The values of red blood cells are significantly lower in postoperative collected autotransfusion blood compared with preoperative collected autologous blood and allogeneic blood (p<0.001). The values of potassium and acid base status were in normal range in postoperatively collected autotransfusion blood. These values in preoperatively collected autologous blood and allogeneic blood were out of normal range; (p<0.001). In addition to reducing the risk of complications that are associated with allogeneic transfusion, postoperative blood salvage may offer benefits including reducing the need for allogeneic blood. Our study confirmed that postoperative collection and transfusion of drainaged blood is simple and safe method that significantly reduce the need for allogeneic transfusion in patients underwent total hip replacement. The blood collected and transfused postoperatively has lower values of red blood cells and normal values of potassium and acid base balance. The transfusion of this blood caused no complications in our patients. 相似文献
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We have characterized the asymmetric effect of Ca2+ on passive K+ permeability in erythrocyte membranes, using inside out and right-side out vesicles. Ca2+, but not Mg2+, can induce an increase in K+ uptake in inside out vesicles. The half-maximal concentration of Ca2+ required to induce the K+ uptake is 0.2 mM, and the permeability increase is not specific for K+. Thus, the Ca2+-induced permeation process in inside out vesicles is changed from that in the energy-depleted intact cell which requires only micromolar concentrations of Ca2+ and is specific for K+. Removal of spectrin had no effect on the vesicle permeability increase due to Ca2+. Studies with show that the vesicle channel opening is mediated by a protein and passage is controlled by sulfhydryl groups; furthermore, the Ca2+-induced vesicle pathway is distinct from the normal channel for passive K+ leak in the absence of Ca2+. The protein is sensitive to its phospholipid environment since removal of easily accessible phospholipid head groups on the cytoplasmic face of the vesicles inhibits the Ca2+-stimulated channel opening. 相似文献
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Cytoplasmic RNA sequences complementary to cloned chick delta-crystallin cDNA show size heterogeneity. 下载免费PDF全文
D J Bower L H Errington N R Wainwright C Sime S Morris R M Clayton 《The Biochemical journal》1982,201(2):339-344
Double-stranded complementary DNA (cDNA) sequences were prepared from day-old chick lens total polysomal RNA and inserted into the unique PstI restriction site of the plasmid pBR322. Colonies containing sequences complementary to abundant lens poly(A)-containing RNA sequences were identified by using lens 32P-labelled cDNA. Some of these clones have been characterized as containing delta-crystallin mRNA coding sequences by genomic DNA blot hybridization and RNA blot hybridizations. Hybridization of labelled DNA from such clones to RNA blots detected four size classes of delta-crystallin RNA sequences, although Southern blots indicated that there are probably only two delta-crystallin genes. 相似文献
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