Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Nat Methods. 2012 Jun 17;9(8):822-4. doi: 10.1038/nmeth.2074.
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.
我们开发了一种使用外膜蛋白酶 T（OmpT）进行受限酶解的方法，用于产生用于基于质谱的蛋白质组学的大肽（平均 >6.3 kDa）。使用这种方法分析预分级的高分子量 HeLa 蛋白质，我们从 1038 种蛋白质中鉴定出 3697 种独特的肽段。我们证明了大 OmpT 肽区分密切相关的蛋白质同工型的能力，并能够检测到许多翻译后修饰。
