7UMB

NanoBRET tracer Tram-bo bound to a KSR2-MEK1 complex

7UMB の概要

• エントリーDOI: 10.2210/pdb7umb/pdb
• 分子名称: Kinase suppressor of Ras 2, Dual specificity mitogen-activated protein kinase kinase 1, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER, ... (4 entities in total)
• 機能のキーワード: tracer, kinase, complex, mek1, ksr2, transferase
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 84924.13

構造登録者

Marsiglia, W.M.,Khan, K.M.,Dar, A.C. (登録日: 2022-04-06, 公開日: 2023-09-27, 最終更新日: 2024-03-20)

主引用文献

Marsiglia, W.M.,Chow, A.,Khan, Z.M.,He, L.,Dar, A.C.
Live-cell target engagement of allosteric MEKi on MEK-RAF/KSR-14-3-3 complexes.
Nat.Chem.Biol., 20:373-381, 2024

PubMed Abstract: The RAS-mitogen-activated protein kinase (MAPK) pathway includes KSR, RAF, MEK and the phospho-regulatory sensor 14-3-3. Specific assemblies among these components drive various diseases and likely dictate efficacy for numerous targeted therapies, including allosteric MEK inhibitors (MEKi). However, directly measuring drug interactions on physiological RAS-MAPK complexes in live cells has been inherently challenging to query and therefore remains poorly understood. Here we present a series of NanoBRET-based assays to quantify direct target engagement of MEKi on MEK1 and higher-order MEK1-bound complexes with ARAF, BRAF, CRAF, KSR1 and KSR2 in the presence and absence of 14-3-3 in living cells. We find distinct MEKi preferences among these complexes that can be compiled to generate inhibitor binding profiles. Further, these assays can report on the influence of the pathogenic BRAF-V600E mutant on MEKi binding. Taken together, these approaches can be used as a platform to screen for compounds intended to target specific complexes in the RAS-MAPK cascade.

PubMed: 37919548
DOI: 10.1038/s41589-023-01454-8

実験手法

X-RAY DIFFRACTION (3.231 Å)