Objective To apply two-dimensional electrophoresis and mass spectrometry in the ovary proteome researchMethods Protein extractions from mouse ovaries were run in IPGphor isoelectric focus system with 11 cm and 24 cm I...Objective To apply two-dimensional electrophoresis and mass spectrometry in the ovary proteome researchMethods Protein extractions from mouse ovaries were run in IPGphor isoelectric focus system with 11 cm and 24 cm IPG strips respectively (pH 3～10, 0.3 mm thick), then the protein spots were identified by mass spectrometry.Results The ovary protein exactions separated by two-dimensional electrophoresis have got high resolution, and identifing protein by mass spectrometry was highly efficient and facilitly. These two techniques should facilitate further investigation of female reproduction proteome research.Conclusion These two rapid high resolutions and efficient techniques have a variety of applications foreground in female reproduction proteome pattern research.展开更多
目的分析牙周炎(periodontitis,PD)与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者唾液中与疾病相关的差异性表达多肽,寻找具有潜在诊断意义的生物标志物,为探讨COPD的早期防治开辟新视角。方法筛选PD患者10例(PD...目的分析牙周炎(periodontitis,PD)与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者唾液中与疾病相关的差异性表达多肽,寻找具有潜在诊断意义的生物标志物,为探讨COPD的早期防治开辟新视角。方法筛选PD患者10例(PD组)、PD伴COPD患者10例(PD伴COPD组)及健康对照人群8例(对照组),收集各组临床资料及唾液样本。应用弱阳离子交换磁珠(weak-cation-exchange magnetic beads,WCX-MB)分离纯化唾液上清样本,应用基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOFMS)系统获取样本生物学信息,进行多肽差异性质谱分析,筛选出组间差异性多肽,并通过液相色谱-质谱/质谱联用技术(liquid chromatography tandem mass spectrometry,LC-MS/MS)对差异性多肽进行鉴定。结果三组样本共检测出77个差异多肽峰,其中PD伴COPD组和PD组间显著性差异多肽峰10个,PD伴COPD组有8个多肽峰(1193.5、1836.2、1735.1、1321.3、1356.8、2086.8、1863.6、2230.9)表达增高,2个多肽峰(1067.3、1124.4)表达减低。其中1193.5、1356.8多肽峰被鉴定为富组蛋白1、颌下腺激素调节蛋白3B及唾液酸性富脯磷蛋白1/2。结论本研究通过WCX-MB和MALDI-TOF-MS分析出PD伴COPD患者唾液中存在与疾病相关的差异性表达多肽,筛选出的差异性表达多肽有望成为早期诊断COPD的辅助指标。展开更多
Alexandrium catenella DH01 is a toxic dinoflagellate species that is able to not only produce paralytic shellfish toxins,but also cause harmful algal blooms along the coast of China.In this study,we presented a new pr...Alexandrium catenella DH01 is a toxic dinoflagellate species that is able to not only produce paralytic shellfish toxins,but also cause harmful algal blooms along the coast of China.In this study,we presented a new protocol for specific labeling and detection of the cell surface proteins(CSPs) of A.catenella DH01 cells using CyDye difference gel electrophoresis(DIGE) fluor minimal dyes.CSPs were identified using two-dimensional gel electrophoresis(2-DE) and MALDI TOF-TOF mass spectrometry(MS).The results showed that the fluorescent cyanine dye Cy3 could specifically label the CSPs of A.catenella DH01,with minimal labeling of intracellular proteins.Among three protein extraction methods evaluated,the Trizol method was the most efficient to extract CSPs with respect to protein spot number and resolution.Forty-one CSPs were separated and identified from A.catenella DH01 by 2-DE,in which 14 were identified in the protein database using MALDI TOF-TOF MS analysis.This work represents the first attempt to investigate the CSPs of A.catenella using the CyDye DIGE fluor dyeing method that provides a potentially important tool for future comprehensive characterization of CSPs and elucidation of the physiological functions of CSPs in dinoflagellates.展开更多
文摘Objective To apply two-dimensional electrophoresis and mass spectrometry in the ovary proteome researchMethods Protein extractions from mouse ovaries were run in IPGphor isoelectric focus system with 11 cm and 24 cm IPG strips respectively (pH 3～10, 0.3 mm thick), then the protein spots were identified by mass spectrometry.Results The ovary protein exactions separated by two-dimensional electrophoresis have got high resolution, and identifing protein by mass spectrometry was highly efficient and facilitly. These two techniques should facilitate further investigation of female reproduction proteome research.Conclusion These two rapid high resolutions and efficient techniques have a variety of applications foreground in female reproduction proteome pattern research.
文摘目的分析牙周炎(periodontitis,PD)与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)患者唾液中与疾病相关的差异性表达多肽,寻找具有潜在诊断意义的生物标志物,为探讨COPD的早期防治开辟新视角。方法筛选PD患者10例(PD组)、PD伴COPD患者10例(PD伴COPD组)及健康对照人群8例(对照组),收集各组临床资料及唾液样本。应用弱阳离子交换磁珠(weak-cation-exchange magnetic beads,WCX-MB)分离纯化唾液上清样本,应用基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOFMS)系统获取样本生物学信息,进行多肽差异性质谱分析,筛选出组间差异性多肽,并通过液相色谱-质谱/质谱联用技术(liquid chromatography tandem mass spectrometry,LC-MS/MS)对差异性多肽进行鉴定。结果三组样本共检测出77个差异多肽峰,其中PD伴COPD组和PD组间显著性差异多肽峰10个,PD伴COPD组有8个多肽峰(1193.5、1836.2、1735.1、1321.3、1356.8、2086.8、1863.6、2230.9)表达增高,2个多肽峰(1067.3、1124.4)表达减低。其中1193.5、1356.8多肽峰被鉴定为富组蛋白1、颌下腺激素调节蛋白3B及唾液酸性富脯磷蛋白1/2。结论本研究通过WCX-MB和MALDI-TOF-MS分析出PD伴COPD患者唾液中存在与疾病相关的差异性表达多肽,筛选出的差异性表达多肽有望成为早期诊断COPD的辅助指标。
基金supported by the National Natural Science Foundation of China (40876059)the National Key Basic Research Program of China(2010CB428703)the Doctoral Fund of Ministry of Education of China(20070384014),the Excellent Group and the Program for New Century Excellent Talents in University to D.Z. Wang
文摘Alexandrium catenella DH01 is a toxic dinoflagellate species that is able to not only produce paralytic shellfish toxins,but also cause harmful algal blooms along the coast of China.In this study,we presented a new protocol for specific labeling and detection of the cell surface proteins(CSPs) of A.catenella DH01 cells using CyDye difference gel electrophoresis(DIGE) fluor minimal dyes.CSPs were identified using two-dimensional gel electrophoresis(2-DE) and MALDI TOF-TOF mass spectrometry(MS).The results showed that the fluorescent cyanine dye Cy3 could specifically label the CSPs of A.catenella DH01,with minimal labeling of intracellular proteins.Among three protein extraction methods evaluated,the Trizol method was the most efficient to extract CSPs with respect to protein spot number and resolution.Forty-one CSPs were separated and identified from A.catenella DH01 by 2-DE,in which 14 were identified in the protein database using MALDI TOF-TOF MS analysis.This work represents the first attempt to investigate the CSPs of A.catenella using the CyDye DIGE fluor dyeing method that provides a potentially important tool for future comprehensive characterization of CSPs and elucidation of the physiological functions of CSPs in dinoflagellates.
