A reference map of a human pituitary adenoma proteome
A reference map of a human pituitary adenoma proteome
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DOI：
10.1002/pmic.200300408
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发表时间：
2003-05-01
期刊：
影响因子：
3.4
通讯作者：
Desiderio, DM
中科院分区：
文献类型：
--
作者：
Zhan, XQ;Desiderio, DM
为了比较垂体腺瘤不同细胞类型的蛋白质组，以实现我们的长期目标，阐明参与垂体腺瘤形成的分子机制，并检测任何肿瘤相关标记物以进行“早期”诊断，本文描述了垂体腺瘤组织蛋白质组的二维凝胶电泳（2-DE）参考图谱。一个垂直的，二维（2-D）聚丙烯酰胺凝胶电泳系统和PDQuest图像分析软件已被用来提供一个高水平的凝胶间的再现性，并准确地阵列在垂体腺瘤组织中表达的每种蛋白质。使用质谱（基质辅助激光解吸/电离-飞行时间MALDI-TOF和液相色谱-电喷雾电离-四极杆-离子阱LC-ESI-Q-IT）和蛋白质数据库来表征2-D凝胶中的每种蛋白质。结果表明，获得了良好的再现性的2-D凝胶图案。四种二维凝胶间匹配斑点的位置偏差在等电聚焦方向为1.95 ± 0.45 mm，在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳方向为1.70 ± 0.53 mm。共CA。2-DE分离出1000个蛋白质点，质谱鉴定出135个蛋白质点，代表111种蛋白质（96个MALDI-TOF点，39个LC-ESI-Q-IT点）。特征性蛋白质包括垂体激素、细胞信号、酶、细胞防御蛋白、细胞结构蛋白、转运蛋白等。这些蛋白质位于细胞质、细胞膜、线粒体、内质网、细胞核、核糖核小体、细胞外组分或分泌于血浆等。这些数据将用于扩展人类垂体的蛋白质组数据库，该数据库可以在网站http://www.utmem.edu上访问。
In order to compare the proteomes from different cell types of pituitary adenomas for our long-term goal to clarify the molecular mechanisms that participate in the formation of pituitary adenoma, and to detect any tumor-related marker for an "early-stage" diagnosis, the two-dimensional gel electrophoresis (2-DE) reference map of a pituitary adenoma tissue proteome is described here. A vertical, two-dimensional (2-D) polyacrylamide gel electrophoresis system and PDQuest image analysis software have been used to provide a high level of between-gel reproducibility and to accurately array each protein expressed in a pituitary adenoma tissue. Mass spectrometry (matrix-assisted laser desorption/ionization-time of flight MALDI-TOF and liquid chromatography-electrospray ionization-quadrupole-ion trap LC-ESI-Q-IT) and protein databases were used to characterize each protein in the 2-D gel. The results demonstrate that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among four 2-D gels was 1.95 +/- 0.45 mm in the isoelectric focusing direction, and 1.70 +/- 0.53 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. A total of ca. 1000 protein spots were separated by 2-DE, and 135 protein spots that represent 111 proteins were characterized with mass spectrometry (96 spots for MALDI-TOF, 39 spots for LC-ESI-Q-IT). The characterized proteins include pituitary hormones, cellular signals, enzymes, cellular-defense proteins, cell-structure proteins, transport proteins, etc. Those proteins were located in the cytoplasmic, cellular membrane, mitochondrial, endoplasmic reticulum, nuclear, ribonucleosome, extracellular fractions, or were secreted in plasma, etc. Those identified proteins contribute to a functional profile of the pituitary adenoma proteome. These data will be used to expand the proteome database of the human pituitary, which can be accessed in the website http://www.utmem.edu /proteomics.