An integrated workflow for crosslinking mass spectrometry
An integrated workflow for crosslinking mass spectrometry
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DOI：
10.15252/msb.20198994
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发表时间：
2019-09-01
影响因子：
9.9
通讯作者：
Rappsilber, Juri
中科院分区：
文献类型：
--
作者：
Mendes, Marta L.;Fischer, Lutz;Rappsilber, Juri
我们提出了一个简洁的工作流程，通过引入顺序消化和交联识别软件XINUMX来增强交联肽的质谱检测。顺序消化通过选择性缩短长胰蛋白酶肽来增强肽检测。我们展示了我们简单的12级分交联多蛋白复合物和细胞裂解物，定量分析和高密度交联的协议，而不需要特定的交联剂功能。这种整体方法揭示了动态蛋白质-蛋白质相互作用位点，这些位点是可接近的，具有基本的功能相关性，因此非常适合于小分子抑制剂的开发。
We present a concise workflow to enhance the mass spectrometric detection of crosslinked peptides by introducing sequential digestion and the crosslink identification software xiSEARCH. Sequential digestion enhances peptide detection by selective shortening of long tryptic peptides. We demonstrate our simple 12-fraction protocol for crosslinked multi-protein complexes and cell lysates, quantitative analysis, and high-density crosslinking, without requiring specific crosslinker features. This overall approach reveals dynamic protein-protein interaction sites, which are accessible, have fundamental functional relevance and are therefore ideally suited for the development of small molecule inhibitors.