Transcription Factor Response Elements on Tip: A Sensitive Approach for Large-Scale Endogenous Transcription Factor Quantitative Identification.
Transcription Factor Response Elements on Tip: A Sensitive Approach for Large-Scale Endogenous Transcription Factor Quantitative Identification.
复制标题
DOI：
10.1021/acs.analchem.6b03150
复制
发表时间：
2016-11
影响因子：
7.4
通讯作者：
Wenhao Shi;Kai Li;Lei Song;Mingwei Liu;Yunzhi Wang;Wanlin Liu;Xia Xia-Xia;Zhaoyu Qin;Bei Zhen-Bei-Zhe
中科院分区：
文献类型：
--
作者：
Wenhao Shi;Kai Li;Lei Song;Mingwei Liu;Yunzhi Wang;Wanlin Liu;Xia Xia-Xia;Zhaoyu Qin;Bei Zhen-Bei-Zhe
在蛋白质组尺度上绘制内源性转录因子（TFs） DNA结合活性的能力将极大地增强我们对各种生物过程的理解。在这里，我们报告了一种高灵敏度，快速，高通量的方法，转录因子响应元件尖端质谱（TOT-MS），允许内源性tf的定量测量。1 μg核提取物的单次质谱检测可在1 h内定量出150个tf。通过TOT结合尖端小尺寸反相液相色谱法，共鉴定了755个tf，与RNA-seq的深度相当。通过研究表皮生长因子（EGF）信号通路中TFs的动态变化，我们进一步证明了TOT-MS的能力。这种方法应该在从有限数量的生物材料阐明TF景观方面找到广泛的应用。
The ability to map endogenous transcription factors (TFs) DNA binding activity at the proteome scale will greatly enhance our understanding of various biological processes. Here we report a highly sensitive, rapid, and high-throughput approach, transcription factor response elements on tip-mass spectrometry (TOT-MS), that allows for quantitative measurement of endogenous TFs. A total of 150 TFs from 1 μg of nuclear extracts can be quantified with single shot mass spectrometry detection in 1 h of machine time. Up to 755 TFs, which is comparable to the depth of RNA-seq, were identified by TOT coupled with on-tip small size reverse-phase liquid chromatography. We further demonstrated the capability of TOT-MS by interrogating the dynamic change of TFs in the epidermal growth factor (EGF) signaling pathway. This approach should find broad applications in elucidating the TF landscape from limited amounts of biological materials.