7AX4

Human TYK2 pseudokinase domain (575-869) in complex with 5-(4-Fluoro-phenyl)-2-ureido-thiophene-3-carboxylic acid amide.

7AX4 の概要

• エントリーDOI: 10.2210/pdb7ax4/pdb
• 分子名称: Non-receptor tyrosine-protein kinase TYK2, 2-(carbamoylamino)-5-(4-fluorophenyl)thiophene-3-carboxamide, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: tyk2 pseudokinase tpca-1, signaling protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 66615.62

構造登録者

Rowland, P. (登録日: 2020-11-09, 公開日: 2021-04-14, 最終更新日: 2024-05-15)

主引用文献

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

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: 33783261
DOI: 10.1177/24725552211002472

実験手法

X-RAY DIFFRACTION (2.12 Å)