Profiling the Dynamic Interfaces of Fluorinated Transcription Complexes for Ligand Discovery and Characterization
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- 作者：William C. Pomerantz ; Ningkun Wang ; Ashley K. Lipinski ; Rurun Wang ; Tomasz Cierpicki ; Anna K. Mapp
- 刊名：ACS Chemical Biology
- 出版年：2012
- 出版时间：August 17, 2012
- 年：2012
- 卷：7
- 期：8
- 页码：1345-1350
- 全文大小：310K
- 年卷期：v.7,no.8(August 17, 2012)
- ISSN：1554-8937
文摘
The conformationally dynamic binding surfaces of transcription complexes present a particular challenge for ligand discovery and characterization. In the case of the KIX domain of the master coactivator CBP/p300, few small molecules have been reported that target its two allosterically regulated binding sites despite the important roles that KIX plays in processes ranging from memory formation to hematopoiesis. Taking advantage of the enrichment of aromatic amino acids at protein interfaces, here we show that the incorporation of six 19F-labeled aromatic side chains within the KIX domain enables recapitulation of the differential binding footprints of three natural activator peptides (MLL, c-Myb, and pKID) in complex with KIX and effectively reports on allosteric changes upon binding using 1D NMR spectroscopy. Additionally, the examination of both the previously described KIX protein鈥損rotein interaction inhibitor Napthol-ASE-phosphate and newly discovered ligand 1-10 rapidly revealed both the binding sites and the affinities of these small molecules. Significantly, the utility of using fluorinated transcription factors for ligand discovery was demonstrated through a fragment screen leading to a new low molecular weight fragment ligand for CBP/p300, 1G7. Aromatic amino acids are enriched at protein鈥揵iomolecule interfaces; therefore, this quantitative and facile approach will be broadly useful for studying dynamic transcription complexes and screening campaigns complementing existing biophysical methods for studying these dynamic interfaces.