Combination of FASP and fully automated 2D-LC-MS/MS allows in-depth proteomic characterization of mouse zymogen granules.
Combination of FASP and fully automated 2D-LC-MS/MS allows in-depth proteomic characterization of mouse zymogen granules.
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DOI：
10.1002/bmc.2805
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发表时间：
2012-09
期刊：
影响因子：
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通讯作者：
Xionghua Sun;Xiaogang Jiang
中科院分区：
文献类型：
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作者：
Xionghua Sun;Xiaogang Jiang
酶原颗粒（ZG）成分在胰腺损伤和疾病中起重要作用。在以前的研究中，蛋白质组学分析与大鼠酶原颗粒分离的二维凝胶电泳或一维SDS-PAGE，然后在凝胶胰蛋白酶消化。为了克服胶内酶切的缺点并对酶原颗粒进行更深入的蛋白质组学分析，本研究结合过滤辅助样品制备方法和全自动2D-LC-MS/MS技术，鉴定了800个ZG蛋白，每个蛋白至少有两个独特的肽段，其中75%以前没有报道过。所鉴定的蛋白质揭示了蛋白质身份和功能的广泛多样性。这是迄今为止最大的ZG蛋白质组数据集，也是第一个小鼠ZG蛋白质组数据集，为进一步研究ZG蛋白质的分子结构和功能奠定了基础。
Zymogen granule (ZG) constituents play important roles in pancreatic injury and disease. In previous studies, proteomic analyses with rat zymogen granules were separated by two-dimensional gel electrophoresis or one-dimensional SDS-PAGE, followed by in-gel tryptic digestion. In order to overcome the disadvantage of in-gel digestion and to carry out further in-depth proteomic analysis of the zymogen granules, in this study, by combining a filter-aided sample preparation method and fully automated 2D-LC-MS/MS technique, 800 ZG proteins were identified with at least two unique peptides for each protein, 75% of which have not been previously reported. The identified proteins revealed broad diversity in protein identity and function. This is the largest dataset of ZG proteome, and also the first dataset of the mouse ZG proteome, which may help elucidate on the molecular architecture of ZGs and their functions.