排序方式: 共有38条查询结果,搜索用时 15 毫秒
1.
Jiang XS Zhou H Zhang L Sheng QH Li SJ Li L Hao P Li YX Xia QC Wu JR Zeng R 《Molecular & cellular proteomics : MCP》2004,3(5):441-455
Four fractions from rat liver (a crude mitochondria (CM) and cytosol (C) fraction obtained with differential centrifugation, a purified mitochondrial (PM) fraction obtained with nycodenz density gradient centrifugation, and a total liver (TL) fraction) were analyzed with two-dimensional liquid chromatography tandem mass spectrometry analysis. A total of 564 rat proteins were identified and were bioinformatically annotated according to their physicochemical characteristics and functions. While most extreme alkaline ribosomal proteins were identified in the TL fraction, the C fraction mainly included neutral enzymes and the PM fraction enriched alkaline proteins and proteins with electron transfer activity or oxygen binding activity. Such characteristics were more apparent in proteins identified only in the TL, C, or PM fraction. The Swiss-Prot annotation and the bioinformatic prediction results proved that the C and PM fractions had enriched cytoplasmic or mitochondrial proteins, respectively. Combination usage of subcellular fractionation with two-dimensional liquid chromatography tandem mass spectrometry was proved to be a high-throughput, sensitive, and effective analytical approach for subcellular proteomics research. Using such a strategy, we have constructed the largest proteome database to date for rat liver (564 rat proteins) and its cytosol (222 rat proteins) and mitochondrial fractions (227 rat proteins). Moreover, the 352 proteins with Swiss-Prot subcellular location annotation in the 564 identified proteins were used as an actual subcellular proteome dataset to evaluate the widely used bioinformatics tools such as PSORT, TargetP, TMHMM, and GRAVY. 相似文献
2.
Jiang XS Tang LY Dai J Zhou H Li SJ Xia QC Wu JR Zeng R 《Molecular & cellular proteomics : MCP》2005,4(7):902-913
We present the first proteomic analysis on the cellular response to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infection. The differential proteomes of Vero E6 cells with and without infection of the SARS-CoV were resolved and quantitated with two-dimensional differential gel electrophoresis followed by ESI-MS/MS identification. Moreover isotope-coded affinity tag technology coupled with two-dimensional LC-MS/MS were also applied to the differential proteins of infected cells. By combining these two complementary strategies, 355 unique proteins were identified and quantitated with 186 of them differentially expressed (at least 1.5-fold quantitative alteration) between infected and uninfected Vero E6 cells. The implication for cellular responses to virus infection was analyzed in depth according to the proteomic results. Thus, the present work provides large scale protein-related information to investigate the mechanism of SARS-CoV infection and pathogenesis. 相似文献
3.
Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma: Identification of potential tumor markers 总被引:12,自引:0,他引:12
Li C Tan YX Zhou H Ding SJ Li SJ Ma DJ Man XB Hong Y Zhang L Li L Xia QC Wu JR Wang HY Zeng R 《Proteomics》2005,5(4):1125-1139
Hepatocellular carcinoma (HCC) is a malignancy of both underdeveloped and developing countries. Proteomes of ten pairs of clinical hepatitis B virus associated HCC tissue samples were obtained by high resolution two-dimensional gel electrophoresis. Comprehensive analyses of proteins associated with B-type HCC were focused on total differentially expressed proteins (> or = two-fold increase or decrease, Student's t-test, p < 0.05) from one pair of samples. Protein identification was done by peptide mass fingerprinting with matrix assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-tandem mass spectrometry. Comparative analyses of proteins associated with B-type HCC included repeat statistics in ten cases. A total of 100 protein spots, corresponding to 80 different gene products, were identified. Proteins whose expression levels were different by more than 2-fold in at least 50% of the cases (five of ten cases) were further analyzed and 45 proteins were selected out as candidates for HCC-associated proteins. Western blotting further validated up-regulated expressions of two candidate proteins in tumor tissues: proliferating cell antigen and stathmin 1. This comprehensive and comparative analyses of proteins associated with B-type HCC could provide useful molecular markers for diagnostics and prognostics and for therapeutic targets. The physiological significance of the differential expressions for several candidate proteins are discussed. 相似文献
4.
利用毛细管电泳分析唾液酸2-氨基吖啶酮(AMAC)衍生物方法, 可在飞摩尔水平分析糖蛋白中唾液酸. 重组人促红细胞生成素(rhu-EPO)、人尿胰蛋白酶抑制剂(hu-UTI)中唾液酸分析结果与文献值符合较好; 而牛α1-酸性糖蛋白(α1-AGP)分析结果与早期文献值相比, 存在一定差异, 并发现该糖蛋白中除含有5-N-乙酰氨基唾液酸(Neu5Ac)外, 还含数量与Neu5Ac相当的5-N-乙醇酰氨基唾液酸(Neu5Gc). 相似文献
5.
人类新型β 3半乳糖基转移酶基因β 3GalT7 的克隆和鉴定 总被引:1,自引:0,他引:1
从人肺 cDNA 文库中克隆到人类β 3- 半乳糖基转移酶家族中的一个新成员β 3GalT7 (AY277592 , EC2.4.1.-) ,并对其进行了鉴定 . 该基因定位在人类染色体 19q13.2. 它包含一个 1 191 bp 的开放阅读框 (ORF). 编码的蛋白质包括一个信号肽和一个半乳糖基转移酶结构域,分子质量和等电点分别为 43.3 ku 和 8.37. 从原核表达中获得的融合蛋白的分子质量和预测相符 . RNA 印迹杂交显示β 3GalT7 在肺、喉和回肠组织中高表达,而在舌、乳房、子宫、睾丸等组织中表达较低 . 此外半定量 RT-PCR 显示β 3GalT7 在人类不同的肿瘤细胞中转录水平有明显差异 . 相似文献
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将小鼠cAMP依赖的蛋白激酶(cAPK)催化亚基α(mCα)分别以成熟、麦芽糖结合蛋白(MBP)融合以及N端连续六个组氨酸(His_6)融合的形式在大肠杆菌中得到了高效表达,且成熟及融合的重组mCα均有明显的蛋白激酶活性,表明蛋白激酶催化核心结构具有相对的独立性。其中His_6-mCα可利用金属离子(Ni ̄(2+))配体亲和层析(Ⅰ-MAC)一步纯化,所得融合蛋白可通过His_6亲合手臂(Tag)固相化于金属离子(Ni ̄(2+))配体亲和树脂上,为进一步利用PhageDisplay多肽库筛选cAPK识别的底物序列和专一性抑制剂打下了基础。 相似文献
8.
Xin-Ling Wang Tao Yu Jin-Song Zhang Qi-Chang Yan Ya-Hong Luo 《Cellular and molecular neurobiology》2009,29(4):549-556
Retinal Müller cells (RMCs) hypertrophy and proliferation play a crucial role in epiretinal membrane formation. This study
was designed to analyze the effects of Fibronectin and specific FAK siRNA in cell adhesion and migration in rat Müller cells.
RMCs were cultured and identified by GFAP, Vimentin, and GLAST mAb, respectively. The cells were planted on dishes coated
with Fibronectin at 0, 1, 5, 10, 50, and 100 μg/ml. The attachment and migration assay was applied to characterize the RMCs–Fibronectin
interactions. Cell lysis and Western blotting were utilized to detect β1-integrin, FAK, and GLAST protein expression. Then the cells were treated with FAK siRNA, non-targeting siRNA, and control
medium. The cell cycle and apoptosis rate was determined by flow cytometry. The attachment, migration, and Western blotting
assay were repeated. These data suggested that almost all the cells expressed GFAP, Vimentin, and GLAST, respectively, which
ensured most of the harvested cells were RMCs. In attachment assay, the A570 values increased significantly with time (F = 1105.439, P < 0.001) and Fibronectin concentration (F = 424.683, P < 0.001). There were significant difference between each Fibronectin concentration in RMCs migration (F = 34.703, P < 0.000). The expression ratio of FAK, β1-integrin, and GLAST elevated significantly as Fibronectin concentration increased (F = 54.755, P < 0.000; F = 119.962, P < 0.000; F = 39.287, P < 0.000). The Fibronectin pretreatment was settled on 50 μg/ml for siRNA inhibition assays. The specific FAK siRNA treatment
significantly increased G0/G1 percentage and apoptosis rate compared with NT siRNA and control group (F = 11.526, P = 0.009; F = 64.772, P < 0.000). The apoptotic rate was significantly suppressed by inhibitors of caspase-8 and 3 (F = 10.500, P = 0.011). The A570 values were significantly suppressed in FAK siRNA groups compared with NT siRNA and control group (F = 154.241, P < 0.000), and the mean migratory cells per view field were significantly decreased (F = 10.906, P = 0.001). FAK and GLAST expression ratio decreased significantly after FAK siRNA treatment (F = 5.315, P = 0.047; F = 5.985, P = 0.042). Take together, FAK is involved in β1-integrin mediated adhesive signaling and play a critical role in regulating Müller cell adhesion, migration, and so far as
to glutamate transportation functions. 相似文献
9.
Human plasma proteome analysis by multidimensional chromatography prefractionation and linear ion trap mass spectrometry identification 总被引:8,自引:0,他引:8
A resurgence of interest in the human plasma proteome has occurred in recent years because it holds great promise of revolution in disease diagnosis and therapeutic monitoring. As one of the most powerful separation techniques, multidimensional liquid chromatography has attracted extensive attention, but most published works have focused on the fractionation of tryptic peptides. In this study, proteins from human plasma were prefractionated by online sequential strong cation exchange chromatography and reversed-phase chromatography. The resulting 30 samples were individually digested by trypsin, and analyzed by capillary reversed-phase liquid chromatography coupled with linear ion trap mass spectrometry. After meeting stringent criteria, a total of 1292 distinct proteins were successfully identified in our work, among which, some proteins known to be present in serum in <10 ng/mL were detected. Compared with other works in published literatures, this analysis offered a more full-scale list of the plasma proteome. Considering our strategy allows high throughput of protein identification in serum, the prefractionation of proteins before MS analysis is a simple and effective method to facilitate human plasma proteome research. 相似文献
10.
Li-Da Chen Jian-Nan Liu Li Lin Zhi Wu Hao Li Yu-Ming Ye Qiao-Zhen Xu Qi-Chang Lin 《PloS one》2015,10(9)