Quantitative Chemical Proteomics Approach To Identify Post-translational Modification-Mediated Protein鈥揚rotein Interactions
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- 作者：Xiang Li ; Emily A. Foley ; Kelly R. Molloy ; Yinyin Li ; Brian T. Chait ; Tarun M. Kapoor
- 刊名：The Journal of the American Chemical Society
- 出版年：2012
- 出版时间：February 1, 2012
- 年：2012
- 卷：134
- 期：4
- 页码：1982-1985
- 全文大小：263K
- 年卷期：v.134,no.4(February 1, 2012)
- ISSN：1520-5126
文摘
Post-translational modifications (PTMs) (e.g., acetylation, methylation, and phosphorylation) play crucial roles in regulating the diverse protein鈥損rotein interactions involved in essentially every cellular process. While significant progress has been made to detect PTMs, profiling protein鈥損rotein interactions mediated by these PTMs remains a challenge. Here, we report a method that combines a photo-cross-linking strategy with stable isotope labeling in cell culture (SILAC)-based quantitative mass spectrometry to identify PTM-dependent protein鈥損rotein interactions. To develop and apply this approach, we focused on trimethylated lysine-4 at the histone H3 N-terminus (H3K4Me3), a PTM linked to actively transcribed gene promoters. Our approach identified proteins previously known to recognize this modification and MORC3 as a new protein that binds H3M4Me3. This study indicates that our cross-linking-assisted and SILAC-based protein identification (CLASPI) approach can be used to profile protein鈥損rotein interactions mediated by PTMs, such as lysine methylation.