2018.4.13 活动预告:新生蛋白的质谱标记技术 (马嫄慧)

报告人：马嫄慧 博士, The Scripps Research Institute

时间：2018-4-13 Friday, 4:30-5:30pm

地点：BRF2 2A03

邀请人：金春玉

Proteomics and Pulse Azidohomoalanine Labeling of Newly Synthesized Proteins: What Are the Potential Applications?

Yuanhui Ma

Cells can adapt to changing environmental conditions by synthesizing newproteins and altering the ensemble of the proteome. Measuring how proteomes respond to perturbations or disease in a narrow time window is crucial to understanding the underlying mechanisms involved. One unbiased classic way in which MS-based analysis can reveal proteome dynamics is through metabolic incorporation with stable isotope labeled amino acids, but it usually takes days to weeks to generate fully-labeled proteomes, and proteins present in low abundance can be missed. In addition, labeling efficiency can be an issue in primary cells and fully differentiated cells. The development of new biorthogonal reactions have provided a new, transformational strategy for labeling proteomes. Azidohomoalanine (AHA), ananalog of methionine, can be accepted by cellular translational machinery and incorporated into newly synthesized proteins (NSPs). AHA-NSPs can be coupled to biotin via CuAAC-mediated click-chemistry and enriched using avidin-based affinity purification. Thus, AHA-containing proteins or peptides can be enriched and efficiently separated from the whole proteome.  In this talk, I will introduce the development of mass spectrometry (MS) based AHA strategies and discuss several potential applications in biological studies.