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41.
Expression profile matrix of Arabidopsis transcription factor genes suggests their putative functions in response to environmental stresses 总被引:39,自引:0,他引:39
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42.
Li S Finley J Liu ZJ Qiu SH Chen H Luan CH Carson M Tsao J Johnson D Lin G Zhao J Thomas W Nagy LA Sha B DeLucas LJ Wang BC Luo M 《The Journal of biological chemistry》2002,277(50):48596-48601
Cytoskeleton-associated proteins (CAPs) are involved in the organization of microtubules and transportation of vesicles and organelles along the cytoskeletal network. A conserved motif, CAP-Gly, has been identified in a number of CAPs, including CLIP-170 and dynactins. The crystal structure of the CAP-Gly domain of Caenorhabditis elegans F53F4.3 protein, solved by single wavelength sulfur-anomalous phasing, revealed a novel protein fold containing three beta-sheets. The most conserved sequence, GKNDG, is located in two consecutive sharp turns on the surface, forming the entrance to a groove. Residues in the groove are highly conserved as measured from the information content of the aligned sequences. The C-terminal tail of another molecule in the crystal is bound in this groove. 相似文献
43.
44.
Processing of viral envelope glycoprotein by the endomannosidase pathway: evaluation of host cell specificity 总被引:4,自引:2,他引:2
Endo-alpha-D-mannosidase is an enzyme involved in N-linked oligosaccharide
processing which through its capacity to cleave the internal linkage
between the glucose-substituted mannose and the remainder of the
polymannose carbohydrate unit can provide an alternate pathway for
achieving deglucosylation and thereby make possible the continued formation
of complex oligosaccharides during a glucosidase blockade. In view of the
important role which has been attributed to glucose on nascent
glycoproteins as a regulator of a number of biological events, we chose to
further define the in vivo action of endomannosidase by focusing on the
well characterized VSV envelope glycoprotein (G protein) which can be
formed by the large array of cell lines susceptible to infection by this
pathogen. Through an assessment of the extent to which the G protein was
converted to an endo-beta-N- acetylglucosaminidase (endo H)-resistant form
during a castanospermine imposed glucosidase blockade, we found that
utilization of the endomannosidase-mediated deglucosylation route was
clearly host cell specific, ranging from greater than 90% in HepG2 and PtK1
cells to complete absence in CHO, MDCK, and MDBK cells, with intermediate
values in BHK, BW5147.3, LLC-PK1, BRL, and NRK cell lines. In some of the
latter group the electrophoretic pattern after endo H treatment suggested
that only one of the two N-linked oligosaccharides of the G protein was
processed by endomannosidase. In the presence of the specific
endomannosidase inhibitor, Glcalpha1-->3(1- deoxy)mannojirimycin, the
conversion of the G protein into an endo H- resistant form was completely
arrested. While the lack of G protein processing by CHO cells was
consistent with the absence of in vitro measured endomannosidase activity
in this cell line, the failure of MDBK and MDCK cells to convert the G
protein into an endo H-resistant form was surprising since these cell lines
have substantial levels of the enzyme. Similarly, we observed that
influenza virus hemagglutinin was not processed in castanospermine-treated
MDCK cells. Our findings suggest that studies which rely on glucosidase
inhibition to explore the function of glucose in controlling such critical
biological phenomena as intracellular movement or quality control should be
carried out in cell lines in which the glycoprotein under study is not a
substrate for endomannosidase action.
相似文献
45.
Chun-Ming Zhang Xing-She Luan Min Xiao Jian Song Lili Lu Xiao Xiao 《Enzyme and microbial technology》2009,44(2):96-100
A bioreactor packed with chitosan immobilized sulfide-oxidase from Streptomyces species LD048 was developed to treat a liquid stream of sulfide. The inoculation system was composed of glass with a 0.7 L working volume and enzyme activity of 2 mmol S g?1 carrier. The sulfide removal efficiency was almost 100% when the volumetric loading was increased up to 3.9 mmol S L?1 h?1 at a space velocity of 18 h?1. The maximal elimination capacity was 22.1 mmol S L?1 h?1 with a space velocity of 72 h?1. When the aeration was increased from 0.05 to 0.1 L min?1, the average removal efficiency improved from 81% to 94%. A removal efficiency of 90% was obtained after 15 days of operation with a load rate of 8.9 mmol S L?1 h?1 and a space velocity of 14.28 h?1. An operational equation based on the ideal plug flow bioreactor and the Michaelis–Menten model predicted the performance of this bioreactor. 相似文献
46.
47.
MiRNAs are small noncoding RNAs that play important roles in various biological processes including tumorigenesis. However, little is known about the expression and function of miR-506 in nasopharyngeal carcinoma (NPC). In this study, we showed that miR-506 was downregulated in nasopharyngeal carcinoma (NPC) cell lines and tissues. Ectopic expression of miR-506 dramatically suppressed cell proliferation, colony formation and invasion. Moreover, we identified the Forkhead box Q1 (FOXQ1) gene as a novel direct target of miR-506. MiR-506 exerts its tumor suppressor function through inhibition of the FOXQ1, which was involved in tumor metastasis and proliferation in various cancers. Furthermore, the expression of FOXQ1 is up-regulated in NPC cell lines and tissues. Taken together, our results indicate that miR-506 functions as a tumor suppressor miRNA in NPC and that its suppressive effects are mediated chiefly by repressing FOXQ1 expression. 相似文献
48.
Yi Liang Jia Cheng Lin Kun Wang Yi Jie Chen Hong Hong Liu Rong Luan Shuai Jiang Tao Che Yong Zhao De Feng Li Da Cheng Wang Lin Guo Hui Sun 《Biochimica et Biophysica Acta (BBA)/General Subjects》2010
Background
We have previously reported a novel fungal galectin Agrocybe aegerita lectin (AAL) with apoptosis-induced activity and nuclear migration activity. The importance of nuclear localization for AAL's apoptosis-induced activity has been established by mutant study. However, the mechanism remains unclear.Methods
We further investigated the mechanism using a previously reported carbohydrate recognition domain (CRD) mutant protein H59Q, which retained its nuclear localization activity but lost most of its apoptotic activity. The cell membrane-binding ability of recombinant AAL (rAAL) and H59Q was analyzed by FACS, and their cellular partners were identified by affinity chromatography and mass spectroscopy. Furthermore, the interaction of AAL and ligand was proved by mammalian two-hybrid and pull down assays. A knockdown assay was used to confirm the role of the ligand.Results
The apoptotic activity of AAL could be blocked by lactose. Mutant H59Q retained comparable cell membrane-binding ability to rAAL. Four cellular binding partners of AAL in HeLa cells were identified: glucose-regulated protein 78 (GRP78); mortality factor 4-like protein 1 (MRG15); elongation factor 2 (EEF2); and heat shock protein 70 (Hsp70). CRD region of AAL was required for the interaction between AAL/mutant AAL and MRG15. MRG15 knockdown increased the cells' resistance to AAL treatment.Conclusion
MRG15 was a nuclear ligand for AAL in HeLa cells. These data implied the existence of a novel nuclear pathway for the antitumor activity of fungal galectin AAL.General significance
These findings provide a novel explanation of AAL bioactivity and contribute to the understanding of mushroom lectins' antitumor activity. 相似文献49.
BtuB is a β‐barrel membrane protein that facilitates transport of cobalamin (vitamin B12) from the extracellular medium across the outer membrane of Escherichia coli. It is thought that binding of B12 to BtuB alters the conformation of its periplasm‐exposed N‐terminal residues (the TonB box), which enables subsequent binding of a TonB protein and leads to eventual uptake of B12 into the cytoplasm. Structural studies determined the location of the B12 binding site at the top of the BtuB's β‐barrel, surrounded by extracellular loops. However, the structure of the loops was found to depend on the method used to obtain the protein crystals, which—among other factors—differed in calcium concentration. Experimentally, calcium concentration was found to modulate the binding of the B12 substrate to BtuB. In this study, we investigate the effect of calcium ions on the conformation of the extracellular loops of BtuB and their possible role in B12 binding. Using all‐atom molecular dynamics, we simulate conformational fluctuations of several X‐ray structures of BtuB in the presence and absence of calcium ions. These simulations demonstrate that calcium ions can stabilize the conformation of loops 3–4, 5–6, and 15–16, and thereby prevent occlusion of the binding site. Furthermore, binding of calcium ions to extracellular loops of BtuB was found to enhance correlated motions in the BtuB structure, which is expected to promote signal transduction. Finally, we characterize conformation dynamics of the TonB box in different X‐ray structures and find an interesting correlation between the stability of the TonB box structure and calcium binding. Proteins 2010. © 2009 Wiley‐Liss, Inc. 相似文献
50.
Human amniotic epithelial cells (hAECs) are potentially one of the key players in tissue engineering due to their easy availability. The aim of the present study was to develop an optimal isolation and transportation technique, as well as to determine the immunophenotype and epithelial gene expression of hAECs. Amnion was mechanically peeled off from the chorion and digested with trypsin-ethylenediaminetetraacetic acid. The isolated hAECs were cultured in medium containing 10 ng/mL epidermal growth factor until P4. The epithelial gene expression, cell surface antigen and protein expression of hAECs were analyzed by quantitative polymerase chain reaction, flow cytometry and immunocytochemistry. hAECs were also cultured in adipogenic, osteogenic and neurogenic induction media. The best cell yield of hAECs was seen in the digestion of 15 pieces of amnion (2 × 2 cm) and isolated 30 min after digestion with trypsin. F12:Dulbecco's modified eagle medium was the best medium for short term storage at 4 °C. hAECs expressed CD9, CD44, CD73 and CD90, and negligibly expressed CD31, CD34, CD45 and CD117. After serial passage, CK3, CK19 and involucrin gene expressions were upregulated, while p63, CK1 and CK14 gene expressions were downregulated. Sustained gene expressions of integrin β1 and CK18 were observed. At initial culture, these cells might have stem-like properties. However, they differentiated after serial passage. Nonetheless, hAECs have epithelial stem cell characteristics and have the potential to differentiate into corneal epithelial cells. Further investigations are still needed to elucidate the mechanism of differentiation involved and to optimize the culture condition for long term in vitro culture. 相似文献