Mediator structure and conformation change

1. Download : Download high-res image (174KB)
2. Download : Download full-size image

     

Molecular docking and pharmacophore model studies of Rho kinase inhibitors

Molecular docking and pharmacophore model approaches were used to characterise the binding features of four different series of Rho kinase (ROCK) inhibitors. Docking simulation of 20 inhibitors with ROCK was performed. The binding conformations and binding affinities of these inhibitors were obtained using AutoDock 4.0 software. The predicted binding affinities correlate well with the activities of these inhibitors (R ² = 0.904). 3D pharmacophore models were generated for ROCK based on highly active inhibitors implemented in Catalyst 4.11 program. The best pharmacophore model consists of one hydrogen bond acceptor feature and two hydrophobic features, and they all seemed to be essential for inhibitors in terms of their binding activities. It is anticipated that the findings reported in this paper may provide very useful information for designing new ROCK inhibitors.     

Resolution and Probabilistic Models of Components in CryoEM Maps of Mature P22 Bacteriophage

CryoEM continues to produce density maps of larger and more complex assemblies with multiple protein components of mixed symmetries. Resolution is not always uniform throughout a cryoEM map, and it can be useful to estimate the resolution in specific molecular components of a large assembly. In this study, we present procedures to 1) estimate the resolution in subcomponents by gold-standard Fourier shell correlation (FSC); 2) validate modeling procedures, particularly at medium resolutions, which can include loop modeling and flexible fitting; and 3) build probabilistic models that combine high-accuracy priors (such as crystallographic structures) with medium-resolution cryoEM densities. As an example, we apply these methods to new cryoEM maps of the mature bacteriophage P22, reconstructed without imposing icosahedral symmetry. Resolution estimates based on gold-standard FSC show the highest resolution in the coat region (7.6 Å), whereas other components are at slightly lower resolutions: portal (9.2 Å), hub (8.5 Å), tailspike (10.9 Å), and needle (10.5 Å). These differences are indicative of inherent structural heterogeneity and/or reconstruction accuracy in different subcomponents of the map. Probabilistic models for these subcomponents provide new insights, to our knowledge, and structural information when taking into account uncertainty given the limitations of the observed density.     

贮藏条件对川硬皮肿腿蜂存活率的影响

在相对湿度为50%~70%的条件下,设置5个温度梯度,将川硬皮肿腿蜂SclerodermasichuanensisXiao雌成蜂分别进行直接贮藏和定期补充15%蜂蜜水后贮藏,并对该蜂在不同条件下的贮藏存活率进行了比较。结果表明,贮藏温度和补充营养对川硬皮肿腿蜂雌成蜂的存活率有显著影响,其中6~15℃下贮藏相同时间的蜂存活率较高,比较适合该蜂的贮藏,而在12℃贮藏相同时间的供试蜂存活率最高,最适该蜂的贮藏。除此之外,在贮藏期间适时添加补充营养也能大大延长川硬皮肿腿蜂雌成蜂的贮藏寿命,但补充营养对该蜂存活率的影响大小与贮藏温度有关。     

Supramolecular aggregation/disaggregation-based molecular sensing: a review focused on investigations from China

Supramolecular aggregation and disaggregation induced by external stimuli can impact the optical or electrical signals of the aggregates/constituting units (receptors). Therefore, manipulating supramolecular aggregation/disaggregation has recently been employed to construct novel and promising photoluminescence (PL)‐based sensing and recognition systems. The sensing systems were capable of substantially enhancing the sensitivity, relying on cooperative interactions occurring in the assembly/disassembly processes (mostly operating in emission turned‐on or emission‐enhanced mode). This review focuses mainly on recent advances in the new emerging PL‐based sensing platforms, based on manipulating the behaviours of supramolecular aggregation/disaggregation, including aggregation‐induced emission (AIE), metallophilic interactions‐related sensing (metallophilic interactions‐induced aggregation/disaggregation), metal coordination polymers‐related sensing, and other sensing systems involving supramolecular aggregation/disaggregation. In particular, those sensing systems developed by scientists in China are summarized and highlighted. Copyright © 2011 John Wiley & Sons, Ltd.     

纵坑切梢小蠹携带云南半帚孢研究

云南半帚孢 (Leptogramphyunnanensis)是纵坑切梢小蠹Tomicuspiniperda重要的共生真菌 ,在纵坑切梢小蠹危害寄主树木过程中发挥着重要作用。研究揭示 ,纵坑切梢小蠹主要通过与受到感染云南半帚孢的韧皮组织的接触携带上云南半帚孢的。纵坑切梢小蠹卵、幼虫和蛹对云南半帚孢的带菌率较高 ,均大于 90 % ,而成虫的带菌率较低。纵坑切梢小蠹的体表和体内均携带有云南半帚孢 ,但体表带菌是纵坑切梢小蠹带菌的主要途径。通过对纵坑切梢小蠹成虫头、足、翅和腹部带菌率的研究发现 ,云南半帚孢在纵坑切梢小蠹各部位的分布大体相同 ,揭示纵坑切梢小蠹没有携带云南半帚孢的特化构造或器官。     

PU.1, a novel capase-3 substrate, partially contributes to chemotherapeutic agents-induced apoptosis in leukemic cells

PU.1 is one of key regulators of hematopoietic cell development, a tightly-regulated lineage-specific process. Here we provide the first evidence that PU.1 protein is cleaved into two fragments of 24 kDa and 16 kDa during apoptosis progression in leukemic cell lines and primary leukemic cells. Further experiments with specific capase-3 inhibitor Z-DEVD-fmk and the in vitro proteolytic system confirmed that PU.1 is a direct target of caspase-3. Using site-directed mutagenesis analyses, the aspartic acid residues at positions 97 and 151 of PU.1 protein were identified as capsase-3 target sites. More intriguingly, the suppression of PU.1 expression by small interfering RNAs (siRNAs) significantly inhibits DNA-damaging agents NSC606985 and etoposide-induced apoptosis in leukemic cells, together with the up-regulated expression of anti-apoptotic bcl-2 gene. These results would provide new insights for understanding the mechanism of PU.1 protein in hematopoiesis and leukemogenesis.     

Characterization of the specific CD4+ T cell response against the F protein during chronic hepatitis C virus infection

Background

The hepatitis C virus (HCV) Alternate Reading Frame Protein (ARFP or F protein) presents a double-frame shift product of the HCV core gene. We and others have previously reported that the specific antibodies against the F protein could be raised in the sera of HCV chronically infected patients. However, the specific CD4⁺ T cell responses against the F protein during HCV infection and the pathological implications remained unclear. In the current study, we screened the MHC class II-presenting epitopes of the F protein through HLA-transgenic mouse models and eventually validated the specific CD4⁺ T cell responses in HCV chronically infected patients.

Methodology

DNA vaccination in HLA-DR1 and-DP4 transgenic mouse models, proliferation assay to test the F protein specific T cell response, genotyping of Chronic HCV patients and testing the F-peptide stimulated T cell response in the peripheral blood mononuclear cell (PBMC) by in vitro expansion and interferon (IFN)- γ intracellular staining.

Principal Findings

At least three peptides within HCV F protein were identified as HLA-DR or HLA-DP4 presenting epitopes by the proliferation assays in mouse models. Further study with human PBMCs evidenced the specific CD4⁺ T cell responses against HCV F protein as well in patients chronically infected with HCV.

Conclusion

The current study provided the evidence for the first time that HCV F protein could elicit specific CD4⁺ T cell response, which may provide an insight into the immunopathogenesis during HCV chronic infection.