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951.
Hansman GS Taylor DW McLellan JS Smith TJ Georgiev I Tame JR Park SY Yamazaki M Gondaira F Miki M Katayama K Murata K Kwong PD 《Journal of virology》2012,86(7):3635-3646
Human noroviruses are genetically and antigenically highly divergent. Monoclonal antibodies raised in mice against one kind of norovirus virus-like particle (VLP), however, were found to have broad recognition. In this study, we present the crystal structure of the antigen-binding fragment (Fab) for one of these broadly reactive monoclonal antibodies, 5B18, in complex with the capsid-protruding domain from a genogroup II genotype 10 (GII.10) norovirus at 3.3-Å resolution and, also, the cryo-electron microscopy structure of the GII.10 VLP at ∼10-Å resolution. The GII.10 VLP structure was more similar in overall architecture to the GV.1 murine norovirus virion than to the prototype GI.1 human norovirus VLP, with the GII.10 protruding domain raised ∼15 Å off the shell domain and rotated ∼40° relative to the GI.1 protruding domain. In the crystal structure, the 5B18 Fab bound to a highly conserved region of the protruding domain. Based on the VLP structure, this region is involved in interactions with other regions of the capsid and is buried in the virus particle. Despite the occluded nature of the recognized epitope in the VLP structure, enzyme-linked immunosorbent assay (ELISA) binding suggested that the 5B18 antibody was able to capture intact VLPs. Together, the results provide evidence that the norovirus particle is capable of extreme conformational flexibility, which may allow for antibody recognition of conserved surfaces that would otherwise be buried on intact particles. 相似文献
952.
Khin-Ohnmar Lwin Hiroki Matsui Tomomi Ban-Tokuda Makoto Kondo Rosalina M. Lapitan Jesus Rommel V. Herrera Arnel N. Del-Barrio Tsutomu Fujihara 《Molecular biology reports》2012,39(12):10769-10774
Comparative analyses of methanogen diversity in the rumen of crossbred buffalo and cattle fed the same diet in the Philippines was performed by cloning the methyl coenzyme M reductase A (mcrA) gene. The cattle and buffalo libraries consisted of 50 clones each. Comparative analysis of the amino acid sequence revealed that these 2 libraries differed significantly (P?<?0.01). The deduced amino acid sequences of the clones were classified into 9 operational taxonomic units (OTUs) in buffalo and 11 OTUs in cattle. Sequence similarity between the clones and known cultured methanogens ranged from 86 to 97?% for buffalo and 84 to 99?% for cattle. Methanobrevibacter species were predominant in buffalo (64?% of the clones), and an unknown mcrA was predominant in cattle (52?% of the clones). A large number of clones with low similarity to cultivated methanogens was observed in both buffalo and cattle, suggesting the presence of an unknown methanogen species in their rumen. 相似文献
953.
Wang W Matsukura M Fujii I Ito K Zhao JE Shinohara M Wang YQ Zhang XM 《Molecular biology reports》2012,39(1):613-620
The aim of this study was to investigate the change of Integrin-linked kinase (ILK) expression of human retinal pigment epithelium
(RPE) cells in response to high glucose, and the effect of targeting ILK with small interference RNA (siRNA) on the high glucose-induced
expression of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1). The ILK mRNA and protein
expression in human RPE cells were analyzed with RT-PCR and western blot after exposure to 5.5, 30, 40, 50 mM glucose, or
5.5 mM glucose + 45.5 mM mannitol for 48 h. The expression of VEGF and ICAM-1 was also determined. Cells were treated with
ILK siRNA, to determine the effect of ILK on VEGF and ICAM-1 expression following treatment with high glucose. High concentrations
of glucose significantly up-regulated ILK mRNA and protein expression, and the ILK expression increased along with the glucose
concentration. The changes of VEGF and ICAM-1 expression were similar to that of ILK expression. Knocking down ILK gene expression
with siRNA inhibited the elevation of VEGF and ICAM-1 induced by high glucose treatment. These results suggested that ILK
was involved in the response of RPE cells to high glucose and may therefore play a role in the pathogenesis of diabetic ophthalmology. 相似文献
954.
Miyazaki T Takenaka T Inoue T Sato M Miyajima Y Nodera M Hanyu M Ohno Y Shibazaki S Suzuki H 《Biological trace element research》2012,145(3):375-381
Zinc deficiency leads to decreased cellular immune responses. The overproduction of nitrogen species derived from inducible
nitric oxide synthase (iNOS), its enzyme, and interleukine-1 beta (IL-1β), and inflammatory cytokine have been implicated
in immune responses. The goal of this study was to investigate the effects of lipopolysaccharide (LPS)-induced changes in
NO metabolites, iNOS, and IL-1β protein expression in the lungs of zinc-deficient rats. Male Sprague–Dawley rats (body weight,
100 g) were divided into two groups and were fed either a zinc-deficient diet (ZnD) or a zinc-containing diet (Cont). After
4 weeks on these diets, rats received a 10-mg/kg dose of LPS injected via the tail vein and were then maintained for an additional
72 h. To determine total NO concentrations in the blood, serum zinc concentration, iNOS protein expression, IL-1β, and iNOS
immunohistochemistry, blood and lung samples were obtained at pre-LPS injection, 5, 24, and 72 h after injection. Total NO
levels were significantly increased at 5, at 24, and at 72 h after LPS injection compared with pre-LPS injection level in
ZnD group; significant changes in total NO levels was elevated at 5 h from at pre-LPS level but not significant changes from
basal level at 24 and 72 h in the control group. Based on western blot analyses and immunohistochemistry, clear bands indicating
iNOS and IL-1β protein expression and iNOS antibody-stained inflammatory cells were detected at 5 and 24 h in the ZnD group
and 5 h in the Cont group, not observed at 24 and 72 h in the control group. These results suggest that zinc deficiency induces
overexpression of iNOS and IL-1β proteins from inflammatory cells around the alveolar blood vessels, resulting in overproduction
of total NO and persisted inflammatory response in the zinc-deficient rat lung. Taken together, overexpression of LPS-induced
iNOS, overproduction of iNOS-derived NO, and overexpression of IL-1β may induce nitrosative and oxidative stresses in the
lung, and these stresses may be involved low immunity of zinc deficiency states. 相似文献
955.
956.
Kayama S Shigemoto N Kuwahara R Onodera M Yokozaki M Ohge H Kato F Hisatsune J Sugai M 《Journal of microbiological methods》2012,88(1):182-184
Klebsiella pneumoniae resistant to almost all ß-lactams except imipenem designated as ISMRK (imipenem-susceptible meropenem-resistant Klebsiella) is emerging in Japan. All ISMRK carries blaIMP-6 which differs from blaIMP-1 by only a single nucleotide at position 640. We devised a rapid detection system of blaIMP-6 by using ARMS PCR. 相似文献
957.
Liu N Matsumoto M Kitagawa K Kotake Y Suzuki S Shirasawa S Nakayama KI Nakanishi M Niida H Kitagawa M 《The EMBO journal》2012,31(10):2365-2377
The tumour suppressor gene product Mig-6 acts as an inhibitor of epidermal growth factor (EGF) signalling. However, its posttranslational modifications and regulatory mechanisms have not been elucidated. Here, we investigated the phosphorylation of human Mig-6 and found that Chk1 phosphorylated Mig-6 in vivo as well as in vitro. Moreover, EGF stimulation promoted phosphorylation of Mig-6 without DNA damage and the phosphorylation was inhibited by depletion of Chk1. EGF also increased Ser280-phosphorylated Chk1, a cytoplasmic-tethering form, via PI3K pathway. Mass spectrometric analyses suggested that Ser 251 of Mig-6 was a major phosphorylation site by Chk1 in vitro and in vivo. Substitution of Ser 251 to alanine increased inhibitory activity of Mig-6 against EGF receptor (EGFR) activation. Moreover, EGF-dependent activation of EGFR and cell growth were inhibited by Chk1 depletion, and were rescued by co-depletion of Mig-6. Our results suggest that Chk1 phosphorylates Mig-6 on Ser 251, resulting in the inhibition of Mig-6, and that Chk1 acts as a positive regulator of EGF signalling. This is a novel function of Chk1. 相似文献
958.
Sakurai T Lee H Kashima M Saito Y Hayashi T Kudome-Takamatsu T Nishimura O Agata K Shibata N 《The International journal of developmental biology》2012,56(1-3):173-182
The growth in size of freshwater planarians in response to nutrient intake is limited by the eventual separation of tail and body fragments in a process called fission. The resulting tail fragment regenerates the entire body as an artificially amputated tail fragment would do, and the body fragment regenerates a tail, resulting in two whole planarians. This regenerative ability is supported by pluripotent somatic stem cells, called neoblasts, which are distributed throughout almost the entire body of the planarian. Neoblasts are the only planarian cells with the ability to continuously proliferate and give rise to all types of cells during regeneration, asexual reproduction, homeostasis, and growth. In order to investigate the molecular characteristics of neoblasts, we conducted an extensive search for neoblast-specific genes using the High Coverage Expression Profiling (HiCEP) method, and tested the function of the resulting candidates by RNAi. Disruption of the expression of one candidate gene, DjP2X-A (Dugesia japonica membrane protein P2X homologue), resulted in a unique phenotype. DjP2X-A RNAi leads to an increase of fission events upon feeding. We confirmed by immunohistochemistry that DjP2X-A is a membrane protein, and elucidated its role in regulating neoblast proliferation, thereby explaining its unique phenotype. We found that DjP2X-A decreases the burst of neoblast proliferation that normally occurs after feeding. We also found that DjP2X-A is required for normal proliferation in starved animals. We propose that DjP2X-A modulates stem cell proliferation in response to the nutritional condition. 相似文献
959.
960.
Goetz R Ohnishi M Ding X Kurosu H Wang L Akiyoshi J Ma J Gai W Sidis Y Pitteloud N Kuro-O M Razzaque MS Mohammadi M 《Molecular and cellular biology》2012,32(10):1944-1954
It has been recently established that Klotho coreceptors associate with fibroblast growth factor (FGF) receptor tyrosine kinases (FGFRs) to enable signaling by endocrine-acting FGFs. However, the molecular interactions leading to FGF-FGFR-Klotho ternary complex formation remain incompletely understood. Here, we show that in contrast to αKlotho, βKlotho binds its cognate endocrine FGF ligand (FGF19 or FGF21) and FGFR independently through two distinct binding sites. FGF19 and FGF21 use their respective C-terminal tails to bind to a common binding site on βKlotho. Importantly, we also show that Klotho coreceptors engage a conserved hydrophobic groove in the immunoglobulin-like domain III (D3) of the "c" splice isoform of FGFR. Intriguingly, this hydrophobic groove is also used by ligands of the paracrine-acting FGF8 subfamily for receptor binding. Based on this binding site overlap, we conclude that while Klotho coreceptors enhance binding affinity of FGFR for endocrine FGFs, they actively suppress binding of FGF8 subfamily ligands to FGFR. 相似文献