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Synthesis,Biochemical Characterization,and Genetic Encoding of a 1,2,4-Triazole Amino Acid as an Acetyllysine Mimic for Bromodomains of the BET Family |
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| Authors: | Dr Sören Kirchgäßner Dr Michael B Braun Natascha Bartlick Dr Cengiz Koç Dr Christopher D Reinkemeier Prof?Dr Edward A Lemke Prof?Dr Thilo Stehle Prof?Dr Dirk Schwarzer |
| Institution: | 1. Interfakultäres Institut für Biochemie, Universität Tübingen, Auf der Morgenstelle 34, 72076 Tübingen, Germany;2. Interfakultäres Institut für Biochemie, Universität Tübingen, Auf der Morgenstelle 34, 72076 Tübingen, Germany
Current address: Department of Infection, Immunity & Cardiovascular Disease, University of Sheffield, The Medical School, Beech Hill Rd, Sheffield, S10 2RX UK;3. Biocenter, Johannes Gutenberg University Mainz, 55128 Mainz, Germany |
| Abstract: | Lysine acetylation is a charge-neutralizing post-translational modification of proteins bound by bromodomains (Brds). A 1,2,4-triazole amino acid (ApmTri) was established as acetyllysine (Kac) mimic recruiting Brds of the BET family in contrast to glutamine commonly used for simulating this modification. Optimization of triazole substituents and side chain spacing allowed BET Brd recruitment to ApmTri-containing peptides with affinities similar to native substrates. Crystal structures of ApmTri-containing peptides in complex with two BET Brds revealed the binding mode which mirrored that of Kac ligands. ApmTri was genetically encoded and recombinant ApmTri-containing proteins co-enriched BRD3(2) from cellular lysates. This interaction was blocked by BET inhibitor JQ1. With genetically encoded ApmTri, biochemistry is now provided with a stable Kac mimic reflecting charge neutralization and Brd recruitment, allowing new investigations into BET proteins in vitro and in vivo. |
| Keywords: | Bromodomain Chemical Biology Genetic Code Expansion Lysine Acetylation Protein Modification |