10.1021/acschembio.8b01020.s005
Maria Reinecke
Maria
Reinecke
Benjamin Ruprecht
Benjamin
Ruprecht
Sandra Poser
Sandra
Poser
Svenja Wiechmann
Svenja
Wiechmann
Mathias Wilhelm
Mathias
Wilhelm
Stephanie Heinzlmeir
Stephanie
Heinzlmeir
Bernhard Kuster
Bernhard
Kuster
Guillaume Médard
Guillaume
Médard
Chemoproteomic Selectivity Profiling of PIKK and PI3K
Kinase Inhibitors
American Chemical Society
2019
novel version
drug targets
Chemoproteomic Selectivity Profiling
Current Kinobeads
ATM inhibitor CP 466722
PIKK
drug discovery
PI 3K inhibitors Omipalisib
chemoproteomic assay
ALK
PI 3K inhibitor action
kinome coverage
cell line
phosphatidylinositol 3- kinases
protein kinases
affinity matrix
NVP-BEZ 235
close-to-physiological conditions
phosphoproteomic experiment
PI 3K inhibitor
PI 3K Kinase Inhibitors Chemical proteomic approaches
broad-selective kinase inhibitors
BGT 226.
PI 3K affinity values
phosphatidylinositol 3- kinase-related kinases
PI 3Ks show
affinity probes
tissue lysates
PI 3Ks
fibrodysplasia ossificans progressiva
2019-03-22 00:00:00
Dataset
https://acs.figshare.com/articles/dataset/Chemoproteomic_Selectivity_Profiling_of_PIKK_and_PI3K_Kinase_Inhibitors/7946567
Chemical
proteomic approaches utilizing immobilized, broad-selective
kinase inhibitors (Kinobeads) have proven valuable for the elucidation
of a compound’s target profile under close-to-physiological
conditions and often revealed potentially synergistic or toxic off-targets.
Current Kinobeads enrich more than 300 native protein kinases from
cell line or tissue lysates but do not systematically cover phosphatidylinositol
3-kinases (PI3Ks) and phosphatidylinositol 3-kinase-related kinases
(PIKKs). Some PIKKs and PI3Ks show aberrant activation in many human
diseases and are indeed validated drug targets. Here, we report the
development of a novel version of Kinobeads that extends kinome coverage
to these proteins. This is achieved by inclusion of two affinity probes
derived from the clinical PI3K/MTOR inhibitors Omipalisib and BGT226.
We demonstrate the utility of the new affinity matrix by the profiling
of 13 clinical and preclinical PIKK/PI3K inhibitors. The large discrepancies
between the PI3K affinity values obtained and reported results from
recombinant assays led us to perform a phosphoproteomic experiment
showing that the chemoproteomic assay is the better approximation
of PI3K inhibitor action <i>in cellulo</i>. The results
further show that NVP-BEZ235 is not a PI3K inhibitor. Surprisingly,
the designated ATM inhibitor CP466722 was found to bind strongly to
ALK2, identifying a new chemotype for drug discovery to treat <i>fibrodysplasia ossificans progressiva</i>.