Transcription Factor Response Elements on Tip: A Sensitive Approach for Large-Scale Endogenous Transcription Factor Quantitative Identification

被引：11

作者：

Shi, Wenhao ^{[1
,2
]}

Li, Kai ^{[2
,6
]}

Song, Lei ^{[1
,2
]}

Liu, Mingwei ^{[2
]}

Wang, Yunzhi ^{[3
,4
]}

Liu, Wanlin ^{[2
]}

Xia, Xia ^{[2
]}

Qin, Zhaoyu ^{[3
,4
]}

Zhen, Bei ^{[2
]}

Wang, Yi ^{[5
]}

He, Fuchu ^{[1
,2
,3
,4
]}

Qin, Jun ^{[2
,3
,4
,5
]}

Ding, Chen ^{[2
,3
,4
]}

机构：

[1] Tsinghua Univ, Sch Life Sci, Beijing 100084, Peoples R China

[2] PHOENIX Ctr, Natl Ctr Prot Sci, Beijing Inst Radiat Med, State Key Lab Prote,Beijing Proteome Res Ctr, Beijing 102206, Peoples R China

[3] Fudan Univ, Sch Life Sci, Inst Biomed Sci, State Key Lab Genet Engn, Shanghai 200032, Peoples R China

[4] Fudan Univ, Sch Life Sci, Inst Biomed Sci, Collaborat Innovat Ctr Genet & Dev, Shanghai 200032, Peoples R China

[5] Baylor Coll Med, Dept Mol & Cellular Biol, Verna & Marrs McLean Dept Biochem & Mol Biol, Alkek Ctr Mol Discovery, Houston, TX 77030 USA

[6] North China Univ Sci & Technol, Sch Basic Med Sci, Dept Pathogeny Biol, Tangshan 063009, Hebei, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2016年 / 88卷 / 24期

基金：

中国国家自然科学基金;

关键词：

PROTEIN-DNA INTERACTIONS; EGF; EXPRESSION;

D O I：

10.1021/acs.analchem.6b03150

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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 mu g 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.

引用

页码：11990 / 11994

页数：5

共 17 条

[1] An Unconventional KITENIN/ErbB4-Mediated Downstream Signal of EGF Upregulates c-Jun and the Invasiveness of Colorectal Cancer Cells [J].