A protease for 'middle-down' proteomics.

A protease for 'middle-down' proteomics.
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DOI:
10.1038/nmeth.2074
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发表时间:
2012-06-17
期刊:
影响因子:
48
通讯作者:
Kelleher, Neil L.
Kelleher, Neil L.
中科院分区:
生物学1区
文献类型:
--
作者:
Wu, Cong;Tran, John C.;Zamdborg, Leonid;Durbin, Kenneth R.;Li, Mingxi;Ahlf, Dorothy R.;Early, Bryan P.;Thomas, Paul M.;Sweedler, Jonathan V.;Kelleher, Neil L.

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我们开发了一种使用外膜蛋白酶T(OmpT)进行限制性酶促蛋白水解以产生大肽(平均> 6.3kDa)用于基于质谱的蛋白质组学的方法。使用这种方法来分析预分级的高质量HeLa蛋白质,我们从1,038种蛋白质中鉴定出3,697种独特的肽。我们证明了大OmpT肽区分密切相关的蛋白质亚型,并使许多翻译后修饰的检测能力。
We developed a method for restricted enzymatic proteolysis using the outer membrane protease T (OmpT) to produce large peptides (> 6.3 kDa on average) for mass spectrometry-based proteomics. Using this approach to analyze prefractionated high-mass HeLa proteins we identified 3,697 unique peptides from 1,038 proteins. We demonstrated the ability of large OmpT peptides to differentiate closely related protein isoforms and to enable the detection of many post-translational modifications.
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