7AX4
Human TYK2 pseudokinase domain (575-869) in complex with 5-(4-Fluoro-phenyl)-2-ureido-thiophene-3-carboxylic acid amide.
Summary for 7AX4
| Entry DOI | 10.2210/pdb7ax4/pdb |
| Descriptor | Non-receptor tyrosine-protein kinase TYK2, 2-(carbamoylamino)-5-(4-fluorophenyl)thiophene-3-carboxamide, CALCIUM ION, ... (4 entities in total) |
| Functional Keywords | tyk2 pseudokinase tpca-1, signaling protein |
| Biological source | Homo sapiens (Human) |
| Total number of polymer chains | 2 |
| Total formula weight | 66615.62 |
| Authors | Rowland, P. (deposition date: 2020-11-09, release date: 2021-04-14, Last modification date: 2024-05-15) |
| Primary citation | Greenhough, L.A.,Clarke, G.,Phillipou, A.N.,Mazani, F.,Karamshi, B.,Rowe, S.,Rowland, P.,Messenger, C.,Haslam, C.P.,Bingham, R.P.,Craggs, P.D. Reducing False Positives through the Application of Fluorescence Lifetime Technology: A Comparative Study Using TYK2 Kinase as a Model System. SLAS Discov, 26:663-675, 2021 Cited by PubMed Abstract: The predominant assay detection methodologies used for enzyme inhibitor identification during early-stage drug discovery are fluorescence-based. Each fluorophore has a characteristic fluorescence decay, known as the fluorescence lifetime, that occurs throughout a nanosecond-to-millisecond timescale. The measurement of fluorescence lifetime as a reporter for biological activity is less common than fluorescence intensity, even though the latter has numerous issues that can lead to false-positive readouts. The confirmation of hit compounds as true inhibitors requires additional assays, cost, and time to progress from hit identification to lead drug-candidate optimization. To explore whether the use of fluorescence lifetime technology (FLT) can offer comparable benefits to label-free-based approaches such as RapidFire mass spectroscopy (RF-MS) and a superior readout compared to time-resolved fluorescence resonance energy transfer (TR-FRET), three equivalent assays were developed against the clinically validated tyrosine kinase 2 (TYK2) and screened against annotated compound sets. FLT provided a marked decrease in the number of false-positive hits when compared to TR-FRET. Further cellular screening confirmed that a number of potential inhibitors directly interacted with TYK2 and inhibited the downstream phosphorylation of the signal transducer and activator of transcription 4 protein (STAT4). PubMed: 33783261DOI: 10.1177/24725552211002472 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.12 Å) |
Structure validation
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