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A proximity-induced covalent fluorescent probe for selective detection of bromodomain 4

Chinese Chemical Letters 2018年第7期29卷 1147-1150页

作者： Ling Feng Mohit Chhabra Wing Ho So Qing Zhu Jiang Xia Hongyan Sun Department of Chemistry and Center of Super-Diamond and Advanced Films (COSDAF) City University of Hong Kong Key Laboratory of Biochip Technology Biotech and Health Centre Shenzhen Research Institute of City University of Hong Kong Department of Chemistry The Chinese University of Hong Kong College of Biotechnology and bioengineering Zhejiang University of Technology

Lysine acetylation is one of the most prevalent and important posttranslational modifications(PTMs) in proteins. The process can be recognized by bromodomains(BRDs), which are a class of proteininteraction modules involved in chromatin remodeling and transcriptional activation. The development of BRD fluorescent probes will be useful for monitoring the activity of BRDs in living cells as well as aiding inhibitor development. Herein we designed a peptide-based probe based on the proximity-induced protein conjugation reaction. The peptide-based probe is capable of covalently and selectively reacting with the unique cysteine residue in the bromodomain through proximity effect. Our experimental data showed that the probe displayed noticeable fluorescence response upon addition of BRD4(1). In-gel fluorescence scanning demonstrated that BRD4(1) can be covalently labelled by the probe. Moreover, the probe was shown to selectively detect BRD4(1) over other proteins. We envision that the probe developed in this study will provide a useful tool to further investigate the biological roles of BRDs.

关键词： peptide-based probe Bromodomain Fluorescent probe Proximity effect peptides

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