Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-d...Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorp-tion/ionization time of flight mass spectrometry (MALDI-TOF-MS). The results showed that well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained under the condition of 0.75-mg protein-load. The average deviation of spot position was 0.733±0.101 mm in IEF direction, and 0.925±0.207 mm in SDS-PAGE direction. For tumor tissue, a total of 1241±88 spots were detected, 987±65 spots were matched with an average matching rate of 79.5%. For control, a total of 1190±72 spots were detected, and 875±48 spots were matched with an average matching rate of 73.5%. A total of 864±34 spots were matched between tumors and controls. Forty-three differential proteins were characterized: some proteins were related to oncogenes, and others involved in the regulation of cell cycle and signal transduc-tion. It is suggested that the differential proteomic approach is valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis. These data will be used to establish human lung cancer proteome database to further study human lung squamous carcinoma.展开更多
用焦锑酸钾沉淀法对45℃和28℃下黄瓜(新泰密刺)授粉后柱头及子房中C a2+分布进行了电镜观察,并采用酶联免疫吸附测定法(EL ISA)测定了叶片中的ABA含量及用等电聚焦聚丙烯酰胺双向电泳(IEF-SDS PAGE)方法对其蛋白合成的变化进行了研究....用焦锑酸钾沉淀法对45℃和28℃下黄瓜(新泰密刺)授粉后柱头及子房中C a2+分布进行了电镜观察,并采用酶联免疫吸附测定法(EL ISA)测定了叶片中的ABA含量及用等电聚焦聚丙烯酰胺双向电泳(IEF-SDS PAGE)方法对其蛋白合成的变化进行了研究.结果表明,28℃时柱头中的C a2+主要分布在细胞间隙中,经45℃处理后细胞间隙和胞内C a2+水平均显著升高,胞内外C a2+浓度梯度逐步丧失;45℃处理1 h后柱头和子房中内源ABA含量显著提高;高温处理后柱头和子房中产生了一些新的蛋白质,如柱头中的(MW26.0 kD,P I 5.4)、(MW90.0 kD,P I5.1)、(MW54.0 kD,P I 4.6)、(MW41.0 kD,P I 6.2),子房中的(MW90.0 kD,P I5.2)、(MW61.0 kD,P I 5.4)、(MW48.0 kD,P I 6.4)、(MW40.5 kD,P I 4.9);另有一些蛋白质与常温相比表达量大幅上调,如柱头中的(MW67.0kD,P I 5.8)、(MW56.5 kD,P I 5.8),子房中的(MW70.0 kD,P I 5.7)、(MW57.0 kD,P I 5.7).上述结果表明,C a2+和ABA信号系统均参与了授粉后黄瓜雌性器官对高温胁迫的反应调节并最终导致基因表达发生变化.展开更多
Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments...Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments of proteomics at the levels of individual plants, tissues, organs and organells, as well as its applications in the area of plant genetic diversity, mutant characterization, and plant physiology, etc are reviewed. At last, the challenge and prospect of proteomics are discussed.展开更多
采用双向凝胶电泳技术优化分离蓝斑背肛海兔(Notarcus leachii cirrosusStimpson,NLCS)口腔神经节(Buccal Ganglion,BG)蛋白质组,并获得约300个蛋白质斑点.用组合基质辅助激光解吸电离化飞行时间(MALD I-TOF)质谱技术和胶内酶解技术测...采用双向凝胶电泳技术优化分离蓝斑背肛海兔(Notarcus leachii cirrosusStimpson,NLCS)口腔神经节(Buccal Ganglion,BG)蛋白质组,并获得约300个蛋白质斑点.用组合基质辅助激光解吸电离化飞行时间(MALD I-TOF)质谱技术和胶内酶解技术测定BG蛋白质组中的96个蛋白质斑点的肽指纹(Peptide m ass fin-gerprint,PMF)图谱.经数据库检索与比对后,发现96种蛋白质中仅有4种蛋白质可获得较高的匹配率,它们分别是微管蛋白(Tubu lin)、肌动蛋白(Actin)和两个1,5-二磷酸核酮糖-羧化酶/加氧酶(R ibu lose-1,5-b i-sphosphate carboxylase/oxygenase,R ibu lose),均属于神经细胞骨架蛋白质;同时还发现一种交配信号肽前体(Peptide m ating pheromone precursor).利用LOC trees软件和分类法对56种蛋白质进行亚细胞定位与分类.展开更多
Two-dimensional polyacrylamide gel assisted laser desorption/ionization electrophoresis （2D-PAGE） and matrixtandem time-of-flight mass spectrometry （MALDI-TOF/TOF-MS）, incorporated with online database searching,...Two-dimensional polyacrylamide gel assisted laser desorption/ionization electrophoresis （2D-PAGE） and matrixtandem time-of-flight mass spectrometry （MALDI-TOF/TOF-MS）, incorporated with online database searching, were performed to investigate differential proteins of breast cancer and adjacent normal breast tissues. Considering that serum albumin is abundantly presented in normal control samples, 15 differential spots detected in 11 out of 12 （91.7%） breast cancer samples were identified by online SIENA-2DPAGE database searching and MALDI-TOF/TOF-MS analysis. The results indicate that pathological changes of breast cancer are concerned with augmentation of substance metabolism, promotion of proteolytic activity, decline of activity of some inhibitors of enzymes, and so on. Some important proteins involved in the pathological process of breast cancer with changed expression may be useful biomarkers, such as alpha-l-antitrypsin, EF- 1-beta, cathepsin D, TCTP, SMT3A, RPS12, and PSMA1, among which SMT3A, RPSl2, and PSMA1 were first reported for breast cancer in this study.展开更多
基金This work was supported by a grant from National 973 Project (2001CB5102) for Outstanding Scholars of New Era from the Ministry of Education of China (2002-48)+1 种基金 National Natural Science Foundation of China (30000028) the key research program from Sc
文摘Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorp-tion/ionization time of flight mass spectrometry (MALDI-TOF-MS). The results showed that well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained under the condition of 0.75-mg protein-load. The average deviation of spot position was 0.733±0.101 mm in IEF direction, and 0.925±0.207 mm in SDS-PAGE direction. For tumor tissue, a total of 1241±88 spots were detected, 987±65 spots were matched with an average matching rate of 79.5%. For control, a total of 1190±72 spots were detected, and 875±48 spots were matched with an average matching rate of 73.5%. A total of 864±34 spots were matched between tumors and controls. Forty-three differential proteins were characterized: some proteins were related to oncogenes, and others involved in the regulation of cell cycle and signal transduc-tion. It is suggested that the differential proteomic approach is valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis. These data will be used to establish human lung cancer proteome database to further study human lung squamous carcinoma.
文摘用焦锑酸钾沉淀法对45℃和28℃下黄瓜(新泰密刺)授粉后柱头及子房中C a2+分布进行了电镜观察,并采用酶联免疫吸附测定法(EL ISA)测定了叶片中的ABA含量及用等电聚焦聚丙烯酰胺双向电泳(IEF-SDS PAGE)方法对其蛋白合成的变化进行了研究.结果表明,28℃时柱头中的C a2+主要分布在细胞间隙中,经45℃处理后细胞间隙和胞内C a2+水平均显著升高,胞内外C a2+浓度梯度逐步丧失;45℃处理1 h后柱头和子房中内源ABA含量显著提高;高温处理后柱头和子房中产生了一些新的蛋白质,如柱头中的(MW26.0 kD,P I 5.4)、(MW90.0 kD,P I5.1)、(MW54.0 kD,P I 4.6)、(MW41.0 kD,P I 6.2),子房中的(MW90.0 kD,P I5.2)、(MW61.0 kD,P I 5.4)、(MW48.0 kD,P I 6.4)、(MW40.5 kD,P I 4.9);另有一些蛋白质与常温相比表达量大幅上调,如柱头中的(MW67.0kD,P I 5.8)、(MW56.5 kD,P I 5.8),子房中的(MW70.0 kD,P I 5.7)、(MW57.0 kD,P I 5.7).上述结果表明,C a2+和ABA信号系统均参与了授粉后黄瓜雌性器官对高温胁迫的反应调节并最终导致基因表达发生变化.
文摘Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments of proteomics at the levels of individual plants, tissues, organs and organells, as well as its applications in the area of plant genetic diversity, mutant characterization, and plant physiology, etc are reviewed. At last, the challenge and prospect of proteomics are discussed.
文摘采用双向凝胶电泳技术优化分离蓝斑背肛海兔(Notarcus leachii cirrosusStimpson,NLCS)口腔神经节(Buccal Ganglion,BG)蛋白质组,并获得约300个蛋白质斑点.用组合基质辅助激光解吸电离化飞行时间(MALD I-TOF)质谱技术和胶内酶解技术测定BG蛋白质组中的96个蛋白质斑点的肽指纹(Peptide m ass fin-gerprint,PMF)图谱.经数据库检索与比对后,发现96种蛋白质中仅有4种蛋白质可获得较高的匹配率,它们分别是微管蛋白(Tubu lin)、肌动蛋白(Actin)和两个1,5-二磷酸核酮糖-羧化酶/加氧酶(R ibu lose-1,5-b i-sphosphate carboxylase/oxygenase,R ibu lose),均属于神经细胞骨架蛋白质;同时还发现一种交配信号肽前体(Peptide m ating pheromone precursor).利用LOC trees软件和分类法对56种蛋白质进行亚细胞定位与分类.
文摘Two-dimensional polyacrylamide gel assisted laser desorption/ionization electrophoresis （2D-PAGE） and matrixtandem time-of-flight mass spectrometry （MALDI-TOF/TOF-MS）, incorporated with online database searching, were performed to investigate differential proteins of breast cancer and adjacent normal breast tissues. Considering that serum albumin is abundantly presented in normal control samples, 15 differential spots detected in 11 out of 12 （91.7%） breast cancer samples were identified by online SIENA-2DPAGE database searching and MALDI-TOF/TOF-MS analysis. The results indicate that pathological changes of breast cancer are concerned with augmentation of substance metabolism, promotion of proteolytic activity, decline of activity of some inhibitors of enzymes, and so on. Some important proteins involved in the pathological process of breast cancer with changed expression may be useful biomarkers, such as alpha-l-antitrypsin, EF- 1-beta, cathepsin D, TCTP, SMT3A, RPS12, and PSMA1, among which SMT3A, RPSl2, and PSMA1 were first reported for breast cancer in this study.
