5LCJ

In-Gel Activity-Based Protein Profiling of a Clickable Covalent Erk 1/2 Inhibitor

5LCJ の概要

• エントリーDOI: 10.2210/pdb5lcj/pdb
• 分子名称: Mitogen-activated protein kinase 1, [(1~{R},4~{Z})-cyclooct-4-en-1-yl] ~{N}-[4-[4-[[5-chloranyl-4-[[2-(propanoylamino)phenyl]amino]pyrimidin-2-yl]amino]pyridin-2-yl]but-3-ynyl]carbamate, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: erk1/2 covalent inhibitor, transferase
• 由来する生物種: Homo sapiens (Human)
• 細胞内の位置: Cytoplasm, cytoskeleton, spindle : P28482
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 43332.15

構造登録者

O'Reilly, M.,Wright, D. (登録日: 2016-06-22, 公開日: 2016-07-20, 最終更新日: 2024-11-13)

主引用文献

Lebraud, H.,Wright, D.J.,East, C.E.,Holding, F.P.,O'Reilly, M.,Heightman, T.D.
In-gel activity-based protein profiling of a clickable covalent ERK1/2 inhibitor.
Mol Biosyst, 12:2867-2874, 2016

PubMed Abstract: In-gel activity-based protein profiling (ABPP) offers rapid assessment of the proteome-wide selectivity and target engagement of a chemical tool. Here we demonstrate the use of the inverse electron demand Diels Alder (IEDDA) click reaction for in-gel ABPP by evaluating the selectivity profile and target engagement of a covalent ERK1/2 probe tagged with a trans-cyclooctene group. The chemical probe was shown to bind covalently to Cys166 of ERK2 using protein MS and X-ray crystallography, and displayed submicromolar GI50s in A375 and HCT116 cells. In both cell lines, the probe demonstrated target engagement and a good selectivity profile at low concentrations, which was lost at higher concentrations. The IEDDA cycloaddition enabled fast and quantitative fluorescent tagging for readout with a high background-to-noise ratio and thereby provides a promising alternative to the commonly used copper catalysed alkyne-azide cycloaddition.

PubMed: 27385078
DOI: 10.1039/c6mb00367b

実験手法

X-RAY DIFFRACTION (1.78 Å)