5U8L

Crystal structure of EGFR kinase domain in complex with a sulfonyl fluoride probe XO44

5U8L の概要

• エントリーDOI: 10.2210/pdb5u8l/pdb
• 分子名称: Epidermal growth factor receptor, 4-[(4-{4-[(3-cyclopropyl-1H-pyrazol-5-yl)amino]-6-[(prop-2-yn-1-yl)carbamoyl]pyrimidin-2-yl}piperazin-1-yl)methyl]benzene-1-sulfonyl fluoride, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: kinase covalent probe covalent inhibitor complex, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Homo sapiens (Human)
• 細胞内の位置: Cell membrane; Single-pass type I membrane protein. Isoform 2: Secreted: P00533
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 38307.21

構造登録者

Gajiwala, K.S.,Ferre, R. (登録日: 2016-12-14, 公開日: 2017-01-25, 最終更新日: 2024-10-30)

主引用文献

Zhao, Q.,Ouyang, X.,Wan, X.,Gajiwala, K.S.,Kath, J.C.,Jones, L.H.,Burlingame, A.L.,Taunton, J.
Broad-Spectrum Kinase Profiling in Live Cells with Lysine-Targeted Sulfonyl Fluoride Probes.
J. Am. Chem. Soc., 139:680-685, 2017

PubMed Abstract: Protein kinases comprise a large family of structurally related enzymes. A major goal in kinase-inhibitor development is to selectively engage the desired kinase while avoiding myriad off-target kinases. However, quantifying inhibitor interactions with multiple endogenous kinases in live cells remains an unmet challenge. Here, we report the design of sulfonyl fluoride probes that covalently label a broad swath of the intracellular kinome with high efficiency. Protein crystallography and mass spectrometry confirmed a chemoselective reaction between the sulfonyl fluoride and a conserved lysine in the ATP binding site. Optimized probe 2 (XO44) covalently modified up to 133 endogenous kinases, efficiently competing with high intracellular concentrations of ATP. We employed probe 2 and label-free mass spectrometry to quantify intracellular kinase engagement by the approved drug, dasatinib. The data revealed saturable dasatinib binding to a small subset of kinase targets at clinically relevant concentrations, highlighting the utility of lysine-targeted sulfonyl fluoride probes in demanding chemoproteomic applications.

PubMed: 28051857
DOI: 10.1021/jacs.6b08536

実験手法

X-RAY DIFFRACTION (1.6 Å)