175885-18-4Relevant articles and documents
Dichloromaleimide (diCMI): A small and fluorogenic reactive group for use in affinity labeling
Chiba, Kosuke,Hashimoto, Yuichi,Yamaguchi, Takao
, p. 1647 - 1653 (2016)
Chemical probes comprising a ligand moiety, a reactive group (e.g. epoxide, haloacetyl or photoreactive group) and a tag unit (e.g. fluorophore or radioisotope) are widely used in affinity labeling to identify the target proteins of bioactive molecules. However, design and synthesis of highly functionalized chemical probes are often time-consuming. In this paper, we propose a simple design strategy for chemical probes bearing a small 2,3-dichloromaleimide (diCMI) unit, which serves as a combined reactive group and tag unit by reacting with a nucleophilic lysine residue near the ligand-binding site of the target protein to generate the 2-amino-3-chloromaleimide fluorophore. Model ligand-protein experiments confirmed that the diCMI unit has suitable reactivity and fluorogenic capability for efficient affinity labeling.
One-Step Synthesis of Photoaffinity Probes for Live-Cell MS-Based Proteomics
Fallon, David J.,Lehmann, Stephanie,Chung, Chun-wa,Phillipou, Alex,Eberl, Christian,Fantom, Ken G. M.,Zappacosta, Francesca,Patel, Vipulkumar K.,Bantscheff, Marcus,Schofield, Christopher J.,Tomkinson, Nicholas C. O.,Bush, Jacob T.
supporting information, p. 17880 - 17888 (2021/09/16)
We present a one-step Ugi reaction protocol for the expedient synthesis of photoaffinity probes for live-cell MS-based proteomics. The reaction couples an amine affinity function with commonly used photoreactive groups, and a variety of handle functionalities. Using this technology, a series of pan-BET (BET: bromodomain and extra-terminal domain) selective bromodomain photoaffinity probes were obtained by parallel synthesis. Studies on the effects of photoreactive group, linker length and irradiation wavelength on photocrosslinking efficiency provide valuable insights into photoaffinity probe design. Optimal probes were progressed to MS-based proteomics to capture the BET family of proteins from live cells and reveal their potential on- and off-target profiles.
Introducing aldehyde functionality to proteins using ligand-directed affinity labeling
Fung, Yi Man Eva,Huang, Yiran,Li, Xiaoyu,Peng, Jianzhao,Song, Yinan,Xiong, Feng
supporting information, p. 6134 - 6137 (2020/06/10)
Aldehyde is a versatile chemical handle for protein modification. Although many methods have been developed to label proteins with aldehyde, target-specific methods amenable to endogenous proteins are limited. Here, we report a simple affinity probe strategy to introduce aldehydes to native proteins. Notably, the probe contains a latent aldehyde functionality that is only exposed upon target binding, thereby enabling a one-pot labeling procedure.
A Chemical Probe for Protein Crotonylation
Bos, Jeffrey,Muir, Tom W.
supporting information, p. 4757 - 4760 (2018/04/16)
Protein lysine crotonylation has emerged as an important post-translational modification (PTM) in the regulation of gene transcription through epigenetic mechanisms. Here we introduce a chemical probe, based on a water-soluble phosphine warhead, which reacts with the crotonyl modification. We show that this reagent is complementary to antibody-based tools allowing detection of endogenous cellular proteins such as histones carrying the crotonylation PTM. The tool is also used to show that the histone acylation activity of the transcriptional coactivator, p300, can be activated by pre-existing lysine crotonylation through a positive feedback mechanism. This reagent provides a versatile and sensitive probe for the analysis of this PTM.
