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71.
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Katoh H Mori Y Kambara H Abe T Fukuhara T Morita E Moriishi K Kamitani W Matsuura Y 《Journal of virology》2011,85(21):10976-10988
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that is kept in a zoonotic transmission cycle between pigs and mosquitoes. JEV causes infection of the central nervous system with a high mortality rate in dead-end hosts, including humans. Many studies have suggested that the flavivirus core protein is not only a component of nucleocapsids but also an important pathogenic determinant. In this study, we identified heterogeneous nuclear ribonucleoprotein A2 (hnRNP A2) as a binding partner of the JEV core protein by pulldown purification and mass spectrometry. Reciprocal coimmunoprecipitation analyses in transfected and infected cells confirmed a specific interaction between the JEV core protein and hnRNP A2. Expression of the JEV core protein induced cytoplasmic retention of hnRNP A2 in JEV subgenomic replicon cells. Small interfering RNA (siRNA)-mediated knockdown of hnRNP A2 resulted in a 90% reduction of viral RNA replication in cells infected with JEV, and the reduction was cancelled by the expression of an siRNA-resistant hnRNP A2 mutant. In addition to the core protein, hnRNP A2 also associated with JEV nonstructural protein 5, which has both methyltransferase and RNA-dependent RNA polymerase activities, and with the 5'-untranslated region of the negative-sense JEV RNA. During one-step growth, synthesis of both positive- and negative-strand JEV RNAs was delayed by the knockdown of hnRNP A2. These results suggest that hnRNP A2 plays an important role in the replication of JEV RNA through the interaction with viral proteins and RNA. 相似文献
73.
De novo sequence design of foldable proteins provides a way of investigating principles of protein architecture. We performed fully automated sequence design for a target structure having a three-helix bundle topology and synthesized the designed sequences. Our design principle is different from the conventional approach, in that instead of optimizing interactions within the target structure, we design the global shape of the protein folding funnel. This includes automated implementation of negative design by explicitly requiring higher free energy of the denatured state. The designed sequences do not have significant similarity to those of any natural proteins. The NMR and CD spectroscopic data indicated that one designed sequence has a well-defined three-dimensional structure as well as alpha-helical content consistent with the target. 相似文献
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Masakazu Ishii Rei Shibata Kazuhisa Kondo Takahiro Kambara Yuuki Shimizu Tohru Tanigawa Yasuko K. Bando Masahiro Nishimura Noriyuki Ouchi Toyoaki Murohara 《The Journal of biological chemistry》2014,289(39):27235-27245
Dipeptidyl peptidase-4 inhibitors are known to lower glucose levels and are also beneficial in the management of cardiovascular disease. Here, we investigated whether a dipeptidyl peptidase-4 inhibitor, vildagliptin, modulates endothelial cell network formation and revascularization processes in vitro and in vivo. Treatment with vildagliptin enhanced blood flow recovery and capillary density in the ischemic limbs of wild-type mice, with accompanying increases in phosphorylation of Akt and endothelial nitric-oxide synthase (eNOS). In contrast to wild-type mice, treatment with vildagliptin did not improve blood flow in ischemic muscles of eNOS-deficient mice. Treatment with vildagliptin increased the levels of glucagon-like peptide-1 (GLP-1) and adiponectin, which have protective effects on the vasculature. Both vildagliptin and GLP-1 increased the differentiation of cultured human umbilical vein endothelial cells (HUVECs) into vascular-like structures, although vildagliptin was less effective than GLP-1. GLP-1 and vildagliptin also stimulated the phosphorylation of Akt and eNOS in HUVECs. Pretreatment with a PI3 kinase or NOS inhibitor blocked the stimulatory effects of both vildagliptin and GLP-1 on HUVEC differentiation. Furthermore, treatment with vildagliptin only partially increased the limb flow of ischemic muscle in adiponectin-deficient mice in vivo. GLP-1, but not vildagliptin, significantly increased adiponectin expression in differentiated 3T3-L1 adipocytes in vitro. These data indicate that vildagliptin promotes endothelial cell function via eNOS signaling, an effect that may be mediated by both GLP-1-dependent and GLP-1-independent mechanisms. The beneficial activity of GLP-1 for revascularization may also be partially mediated by its ability to increase adiponectin production. 相似文献
78.
Yumiko Nagai Manabu Inobe Kokichi Kikuchi Toshimitsu Uede 《Microbiology and immunology》1993,37(8):623-632
Rat CD4+ T cells were divided into two distinct subsets by a monoclonal antibody RTH-1 recognizing a unique epitope on rat CD45R. Cellular distribution of OX-22- and RTH-1-defined antigens was the same. However, OX-22 and RTH-1 recognized different epitopes that exist on rat CD45R. The expression of IL-4 gene was detected only in RTH-1low CD4+ T cell subset upon various stimulations. In contrast, the expression of IL-2 and IFN-γ gene varied depending upon the nature of stimuli. The increased cell surface expression of CD44 was detected in RTH-1high CD4+ T cell subset. Conversely the increased expression of CD2 was detected in RTH-1low CD4+ T cell subset. The expression of CD3 and LFA-1 was not significantly different between RTH-1high and RTH-1low subsets. 相似文献
79.
Duk Shin Hidenori Watanabe Hiroyuki Kambara Atsushi Nambu Tadashi Isa Yukio Nishimura Yasuharu Koike 《PloS one》2012,7(10)
Electrocorticography (ECoG) has drawn attention as an effective recording approach for brain-machine interfaces (BMI). Previous studies have succeeded in classifying movement intention and predicting hand trajectories from ECoG. Despite such successes, however, there still remains considerable work for the realization of ECoG-based BMIs as neuroprosthetics. We developed a method to predict multiple muscle activities from ECoG measurements. We also verified that ECoG signals are effective for predicting muscle activities in time varying series when performing sequential movements. ECoG signals were band-pass filtered into separate sensorimotor rhythm bands, z-score normalized, and smoothed with a Gaussian filter. We used sparse linear regression to find the best fit between frequency bands of ECoG and electromyographic activity. The best average correlation coefficient and the normalized root-mean-square error were 0.92±0.06 and 0.06±0.10, respectively, in the flexor digitorum profundus finger muscle. The δ (1.5∼4Hz) and γ2 (50∼90Hz) bands contributed significantly more strongly than other frequency bands (P<0.001). These results demonstrate the feasibility of predicting muscle activity from ECoG signals in an online fashion. 相似文献
80.
Toshimitsu Hoshino Kumiko Kose Yoshihiko Uratani 《Molecular & general genetics : MGG》1990,220(3):461-467
Summary The gene braB, encoding the Na–-coupled carrier for branched-chain amino acids in Pseudomonas aeruginosa PAO, was cloned on cosmid pMMB34. The cosmid clones carrying the braB gene were identified as those that restored growth at low leucine concentration and Na–-dependent leucine transport activity to P. aeruginosa PAO3536 defective in the transport of branched-chain amino acids. Determination of the nucleotide sequence of the DNA fragment shows that the braB gene comprises 1311 bp and encodes a hydrophobic protein of 437 amino acids with a calculated Mr of 45279. The hydropathy profile suggests that there exist in the carrier protein 12 hydrophobic segments long enough to traverse the membrane. The amino acid sequence shows a high degree of homology with thebrnQ product, a branched-chain amino acid carrier of Salmonella typhimurium, while no homology in the nucleotide sequences is found in the braB and brnQ genes. 相似文献