TY - JOUR
T1 - Thermal proteome profiling identifies the membrane-bound purinergic receptor P2X4 as a target of the autophagy inhibitor indophagolin
AU - Carnero Corrales, Marjorie A.
AU - Zinken, Sarah
AU - Konstantinidis, Georgios
AU - Rafehi, Muhammad
AU - Abdelrahman, Aliaa
AU - Wu, Yao-Wen
AU - Janning, Petra
AU - Müller, Christa E.
AU - Laraia, Luca
AU - Waldmann, Herbert
PY - 2021
Y1 - 2021
N2 - Signaling pathways are frequently activated through signal-receiving membrane proteins, and the discovery of small molecules targeting these receptors may yield insights into their biology. However, due to their intrinsic properties, membrane protein targets often cannot be identified by means of established approaches, in particular affinity-based proteomics, calling for the exploration of new methods. Here, we report the identification of indophagolin as representative member of an indoline-based class of autophagy inhibitors through a target-agnostic phenotypic assay. Thermal proteome profiling and subsequent biochemical validation identified the purinergic receptor P2X4 as a target of indophagolin, and subsequent investigations suggest that indophagolin targets further purinergic receptors. These results demonstrate that thermal proteome profiling may enable the de novo identification of membrane-bound receptors as cellular targets of bioactive small molecules.
AB - Signaling pathways are frequently activated through signal-receiving membrane proteins, and the discovery of small molecules targeting these receptors may yield insights into their biology. However, due to their intrinsic properties, membrane protein targets often cannot be identified by means of established approaches, in particular affinity-based proteomics, calling for the exploration of new methods. Here, we report the identification of indophagolin as representative member of an indoline-based class of autophagy inhibitors through a target-agnostic phenotypic assay. Thermal proteome profiling and subsequent biochemical validation identified the purinergic receptor P2X4 as a target of indophagolin, and subsequent investigations suggest that indophagolin targets further purinergic receptors. These results demonstrate that thermal proteome profiling may enable the de novo identification of membrane-bound receptors as cellular targets of bioactive small molecules.
KW - Autophagy
KW - Biological chemistry and chemical biology
KW - Proteomics
KW - Target identification
KW - Thermal proteome profiling
U2 - 10.1016/j.chembiol.2021.02.017
DO - 10.1016/j.chembiol.2021.02.017
M3 - Journal article
C2 - 33725479
SN - 2451-9456
VL - 28
SP - 1750
EP - 1757
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 12
ER -