Sequence-specific capture of protein-DNA complexes for mass spectrometric protein identification.
Sequence-specific capture of protein-DNA complexes for mass spectrometric protein identification.
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DOI：
10.1371/journal.pone.0026217
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Smith LM
中科院分区：
文献类型：
--
作者：
Wu CH;Chen S;Shortreed MR;Kreitinger GM;Yuan Y;Frey BL;Zhang Y;Mirza S;Cirillo LA;Olivier M;Smith LM
The regulation of gene transcription is fundamental to the existence of complex multicellular organisms such as humans. Although it is widely recognized that much of gene regulation is controlled by gene-specific protein-DNA interactions, there presently exists little in the way of tools to identify proteins that interact with the genome at locations of interest. We have developed a novel strategy to address this problem, which we refer to as GENECAPP, for Global ExoNuclease-based Enrichment of Chromatin-Associated Proteins for Proteomics. In this approach, formaldehyde cross-linking is employed to covalently link DNA to its associated proteins; subsequent fragmentation of the DNA, followed by exonuclease digestion, produces a single-stranded region of the DNA that enables sequence-specific hybridization capture of the protein-DNA complex on a solid support. Mass spectrometric (MS) analysis of the captured proteins is then used for their identification and/or quantification. We show here the development and optimization of GENECAPP for an in vitro model system, comprised of the murine insulin-like growth factor-binding protein 1 (IGFBP1) promoter region and FoxO1, a member of the forkhead rhabdomyosarcoma (FoxO) subfamily of transcription factors, which binds specifically to the IGFBP1 promoter. This novel strategy provides a powerful tool for studies of protein-DNA and protein-protein interactions.
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影响因子：
14.9
作者：
Butala M;Busby SJ;Lee DJ
通讯作者：
Lee DJ
影响因子：
7
作者：
Mittler, Gerhard;Butter, Falk;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
14.8
作者：
Lee, Tong Ihn;Johnstone, Sarah E.;Young, Richard A.
通讯作者：
Young, Richard A.
影响因子：
64.8
作者：
Hecht, A;StrahlBolsinger, S;Grunstein, M
通讯作者：
Grunstein, M
DOI：
10.1006/meth.1996.0407
发表时间：
1997-02-01
期刊：
METHODS-A COMPANION TO METHODS IN ENZYMOLOGY
影响因子：
--
作者：
Orlando, V;Strutt, H;Paro, R
通讯作者：
Paro, R