Developing the wool proteome
Developing the wool proteome
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DOI：
10.1016/j.jprot.2010.05.005
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发表时间：
2010-08-05
影响因子：
3.3
通讯作者：
Dyer, Jolon M.
中科院分区：
文献类型：
--
作者：
Clerens, Stefan;Cornellison, Charisa D.;Dyer, Jolon M.
由于缺乏数据库覆盖、蛋白质提取能力差和动态范围问题，羊毛蛋白质组在很大程度上是未知的。然而，研究羊毛物理性质与蛋白质含量之间的相关性，或表征紫外线、热或加工诱导的蛋白质损伤，需要一个可识别的和可识别的蛋白质组。在本研究中，我们通过全面的数据获取、迭代的蛋白质鉴定/验证和序列数据库的并行扩充的策略，实现了前所未有的羊毛蛋白质组鉴定。数据采集包括一系列不同的联用MS技术，包括LC-MS/MS、LC-MALDI、2D-LC-MS/MS和SDS-PAGE LC-MS。利用数据库的迭代搜索和ProteinScape的搜索结果组合，实现了序列数据库中可识别蛋白质的系统扩展。随后对蛋白质鉴定进行了广泛的验证和合理化。总共添加了72个完整的绵羊特有蛋白质序列和30个部分绵羊特有蛋白质序列，鉴定了113个羊毛蛋白质。加强对绵羊特有蛋白质的鉴定和表征将促进所有羊毛纤维蛋白质化学和蛋白质组学的研究。(C)2010爱思唯尔B.V.保留所有权利。
The wool proteome has been largely uncharted due to a lack of database coverage, poor protein extractability and dynamic range issues. Yet, investigating correlations between wool physical properties and protein content, or characterising UV-, heat- or processing-induced protein damage requires the availability of an identifiable and identified proteome.In this study we have achieved unprecedented wool proteome identification through a strategy of comprehensive data acquisition, iterative protein identification/validation and concurrent augmentation of the sequence database. Data acquisition comprised a range of different hyphenated MS techniques including LC-MS/MS, LC-MALDI, 2D-LC-MS/MS and SDS-PAGE LC-MS. Using iterative searching of databases and search result combination using ProteinScape, a systematic expansion of identifiable proteins in the sequence database was achieved. This was followed by extensive validation and rationalisation of the protein identifications. In total, 72 complete and 30 partial ovine-specific protein sequences were added to the database, and 113 wool proteins were identified.Enhanced access to ovine-specific protein identification and characterisation will facilitate all wool fibre protein chemistry and proteomics research. (C) 2010 Elsevier B.V. All rights reserved.