5U8L
Crystal structure of EGFR kinase domain in complex with a sulfonyl fluoride probe XO44
Summary for 5U8L
| Entry DOI | 10.2210/pdb5u8l/pdb |
| Descriptor | 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) |
| Functional Keywords | kinase covalent probe covalent inhibitor complex, transferase-transferase inhibitor complex, transferase/transferase inhibitor |
| Biological source | Homo sapiens (Human) |
| Cellular location | Cell membrane; Single-pass type I membrane protein. Isoform 2: Secreted: P00533 |
| Total number of polymer chains | 1 |
| Total formula weight | 38307.21 |
| Authors | Gajiwala, K.S.,Ferre, R. (deposition date: 2016-12-14, release date: 2017-01-25, Last modification date: 2024-10-30) |
| Primary citation | 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 Cited by 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: 28051857DOI: 10.1021/jacs.6b08536 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.6 Å) |
Structure validation
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