FM-381

Reagent Authentication Certificate:

Chemical Probes Portal FM-381.pdf

Protein target name:

JAK3

Mechanism of action:

Covalent inhibitor

Recommended Concentration for use in cells:

100 nM - 1 uM

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

HUGO ID:

HGNC:6193

Entrez gene ID:

3718

UNIPROT ID:

P52333

Target alias:

Janus kinase 3, JAK-3_HUMAN, JAKL, L-JAK, LJAK, JAK3

Probe Details

Chemical Probes Portal ID:

CPP1521

Probe alias:

BDBM202553

PubChem CID:

122197584

SMILES string:

N(C)(C)C(C(\C#N)=C\C(=CC=C1C2N(C(C(N=2)=C2)=C(C(=N2)N2)C=C2)C(C2)CCCC2)O1)=O

InChi Key:

GJMZWYLOARVASY-NTCAYCPXSA-N

Control compound:

FM-479

Orthogonal probe:

TofacitinibJAK3i

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50(app)

0.154 nM

Potency assay:

Radiometric assay

PDB ID for probe-target interaction (3D structure):

5LWM

Structure-activity relationship:

Yes, see Cell Chem Biol paper

Off target activity

Potency (off target):

IC50(app)

Off target protein and potency:

JAK1: 52 nM, JAK2: 346 nMTYK2: 459 nM, BTK >5 uMITK: 1.8 uM, EGFR >5 uM

Potency assay (off target):

In addition to assays assessing acivity against closely related kinases and those with a Cys comparable to Cys909 found in the gatekeeper position on JAK3 (above), FM-381 was assayed in a panel of 410 protein kinases (ProQinase), with no significant activity detected at 100 nM and weak activity detected at 500 nM.

Probe Selectivity in Vitro:

FM-381 was largely inactive against a panel of bromodomain proteins. The most potent activity was: TAF1 IC50=500 nM.

In cell validation

On target activity in cells

Potency in cells:

EC50

100 nM

Potency assay (cells):

BRET assay in HeLa cells. FM-381 also inhibited IL-12-stimulated phosphorylation of STAT5 in human CD4+ T cells at 100 nM.

Target engagement assay (cells):

Yes, BRET assay in HeLa cells

Off target activity in cells

Potency assay, off target (cells):

In human CD4+ T cells, FM-381 did not inhibit IL-6-stimulated STAT3 phosphorylation at 1 uM.

Primary Reference

Reference (doi):

10.1016/j.chembiol.2016.10.008

Reference (PMID):

27840070

FM-381

Probe Information

Target Details

Janus kinase 3, JAK-3_HUMAN, JAKL, L-JAK, LJAK, JAK3

Probe Details

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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FM-381

Probe Information

Target Details

Janus kinase 3, JAK-3_HUMAN, JAKL, L-JAK, LJAK, JAK3

Probe Details

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?