Photocaged Quinone Methide Crosslinkers for Light‐Controlled Chemical Crosslinking of Protein–Protein and Protein–DNA Complexes. Issue 52 (8th November 2019)

• Record Type: Journal Article
• Authors: Liu, Jun; Cai, Lingchao; Sun, Wei; Cheng, Rujin; Wang, Nanxi; Jin, Ling; Rozovsky, Sharon; Seiple, Ian B.; Wang, Lei
• Is Part Of: Angewandte Chemie international edition. Volume 58:Issue 52(2019)
• Page Start: 18839
• Page End: 18843
• Publication Date: 2019-11-08
• Subjects: crosslinker -- DNA–protein interactions -- photoaffinity labeling -- protein–protein interactions -- quinone methide; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201910135
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Small‐molecule crosslinkers are invaluable for probing biomolecular interactions and for crosslinking mass spectrometry. Existing chemical crosslinkers target only a small selection of amino acids, while conventional photo‐crosslinkers target almost all residues non‐specifically, complicating data analysis. Herein, we report photocaged quinone methide (PQM)‐based crosslinkers that target nine nucleophilic residues through Michael addition, including Gln, Arg, and Asn, which are inaccessible to existing chemical crosslinkers. PQM crosslinkers were used in vitro, in Escherichia coli, and in mammalian cells to crosslink dimeric proteins and endogenous membrane receptors. The heterobifunctional crosslinker NHQM could crosslink proteins to DNA, for which few crosslinkers exist. The photoactivatable reactivity of these crosslinkers and their ability to target multiple amino acids will enhance the use of chemical crosslinking for studies of protein–protein and protein–DNA networks and for structural biology. Abstract : A lightbulb moment : Photocaged quinone methide (PQM) crosslinkers that target nine nucleophilic amino acids, including Gln, Arg, and Asn, have been developed. These can be used in vitro, in Escherichia coli, and in mammalian cells to study protein–protein and protein–DNA interactions.