Shotgun proteomic analysis of microdissected postmortem human pituitary using complementary two-dimensional liquid chromatography coupled with tandem mass spectrometer
Shotgun proteomic analysis of microdissected postmortem human pituitary using complementary two-dimensional liquid chromatography coupled with tandem mass spectrometer
复制标题
使用互补二维液相色谱和串联质谱仪对死后显微解剖的人垂体进行鸟枪法蛋白质组学分析
DOI：
10.1016/j.aca.2010.12.032
复制
发表时间：
2011-03-04
影响因子：
6.2
通讯作者：
Zhu, Jianhong
中科院分区：
文献类型：
--
作者：
Liu, Yingchao;Zhuang, Dongxiao;Zhu, Jianhong
垂体负责多种稳态功能，包括代谢、生长和生殖。蛋白质组分析为垂体蛋白质表达的全面分析提供了一种有效的方法。垂体通常是从死后标本中获得的，这可能会通过蛋白质水解影响蛋白质组谱。本研究的目的是确定死后垂体是否可以用于蛋白质组学分析结合激光捕获显微切割（LCM）。从LCM捕获的催乳素（PRL）细胞的消化肽分离的二维纳米液相色谱（2D-nanoLC/MS）和串联质谱（MS）的特点。所有的MS/MS谱经SEQUEST检索，鉴定出1660个蛋白质。类别分析的蛋白质组揭示了广泛的无偏见的访问细胞组分蛋白质具有不同的功能特性。结果证明了使用2D-nanoLC/MS通过显微切割对有限的蛋白质量进行灵敏的蛋白质组学分析的能力。在肽和蛋白质水平上对所讨论的蛋白质组和来自催乳素瘤对照的蛋白质组进行了详细的比较，表明这两种蛋白质组具有相似的特征。总之，我们的研究结果首次揭示了使用死后人垂体进行蛋白质组学研究的可能性，这对于进一步研究疾病生物标志物鉴定和催乳素瘤肿瘤发生的分子机制具有重要意义。(C)2011爱思唯尔有限公司版权所有。
The pituitary is responsible for multiple homeostatic functions including metabolism, growth and reproduction. Proteome analysis offers an efficient approach for a comprehensive analysis of pituitary protein expression. The pituitary is usually acquired from postmortem specimens, which may potentially affect the proteome profile by proteolysis. The aim of this study was to determine whether the postmortem pituitary could be used in proteomic analysis combining with Laser capture microdissection (LCM). Digested peptides from LCM captured prolactin (PRL) cells were separated by two dimensional-nanoscale liquid chromatography (2D-nanoLC/MS) and characterized by tandem mass spectrometry (MS). All MS/MS spectrums were searched by SEQUEST and a proteorne of 1660 proteins was identified. Category analysis of the proteome revealed an extensive unbiased access to cell component proteins with diverse functional characteristics. The results demonstrated the ability of using 2D-nanoLC/MS to perform sensitive proteomic analysis on limited protein quantities through microdissection. Detailed comparisons between the proteome in question and the one derived from the prolactinoma controls at peptide and protein levels indicated that the two proteomes had similar characters. Overall, our results revealed for the first time the possibility of use of postmortem human pituitary for proteomic research which is important for further studies on disease biomarker identification and molecular mechanisms of prolactinoma tumorigenesis. (C) 2011 Elsevier B.V. All rights reserved.