5LCJ

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

Summary for 5LCJ

• Entry DOI: 10.2210/pdb5lcj/pdb
• Descriptor: 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)
• Functional Keywords: erk1/2 covalent inhibitor, transferase
• Biological source: Homo sapiens (Human)
• Cellular location: Cytoplasm, cytoskeleton, spindle : P28482
• Total number of polymer chains: 1
• Total formula weight: 43332.15

Authors

O'Reilly, M.,Wright, D. (deposition date: 2016-06-22, release date: 2016-07-20, Last modification date: 2024-11-13)

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 (1.78 Å)