5LCK

A Clickable Covalent ERK 1/2 Inhibitor

Summary for 5LCK

• Entry DOI: 10.2210/pdb5lck/pdb
• Descriptor: Mitogen-activated protein kinase 1, SULFATE ION, ~{N}-[2-[[2-[(5-methoxypyridin-3-yl)amino]-5-(trifluoromethyl)pyrimidin-4-yl]amino]phenyl]propanamide, ... (4 entities in total)
• Functional Keywords: erk2 covalent inhibitor, transferase
• Biological source: Homo sapiens (Human)
• Cellular location: Cytoplasm, cytoskeleton, spindle : P28482
• Total number of polymer chains: 1
• Total formula weight: 42413.54

Authors

O'Reilly, M.,Wright, D. (deposition date: 2016-06-22, release date: 2016-07-20, Last modification date: 2024-10-23)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.89 Å)