TY - JOUR
T1 - The Ligand Binding Landscape of Diacylglycerol Kinases
AU - Franks, Caroline E.
AU - Campbell, Sean T.
AU - Purow, Benjamin W.
AU - Harris, Thurl E.
AU - Hsu, Ku Lung
N1 - Funding Information:
We thank Mark M. Ross and Adam Borne for assistance with experiments and manuscript preparation. We thank all members of the Hsu Lab and colleagues for review of the manuscript. This work was supported by the LaunchPad for Diabetes Program funded by the Manning Family Foundation at the University of Virginia (K.-L.H.), University of Virginia Start-up Funds (K.-L.H.), and the NIH ( DA035864 to K.-L.H.; T32 GM007055 to C.E.F.; T32 CA009109 to S.T.C.; DK101946 to T.E.H; CA180699 and CA189524 to B.W.P.).
Publisher Copyright:
© 2017 Elsevier Ltd
PY - 2017/7/20
Y1 - 2017/7/20
N2 - Diacylglycerol kinases (DGKs) are integral components of signal transduction cascades that regulate cell biology through ATP-dependent phosphorylation of the lipid messenger diacylglycerol. Methods for direct evaluation of DGK activity in native biological systems are lacking and needed to study isoform-specific functions of these multidomain lipid kinases. Here, we utilize ATP acyl phosphate activity-based probes and quantitative mass spectrometry to define, for the first time, ATP and small-molecule binding motifs of representative members from all five DGK subtypes. We use chemical proteomics to discover an unusual binding mode for the DGKα inhibitor, ritanserin, including interactions at the atypical C1 domain distinct from the ATP binding region. Unexpectedly, deconstruction of ritanserin yielded a fragment compound that blocks DGKα activity through a conserved binding mode and enhanced selectivity against the kinome. Collectively, our studies illustrate the power of chemical proteomics to profile protein-small molecule interactions of lipid kinases for fragment-based lead discovery.
AB - Diacylglycerol kinases (DGKs) are integral components of signal transduction cascades that regulate cell biology through ATP-dependent phosphorylation of the lipid messenger diacylglycerol. Methods for direct evaluation of DGK activity in native biological systems are lacking and needed to study isoform-specific functions of these multidomain lipid kinases. Here, we utilize ATP acyl phosphate activity-based probes and quantitative mass spectrometry to define, for the first time, ATP and small-molecule binding motifs of representative members from all five DGK subtypes. We use chemical proteomics to discover an unusual binding mode for the DGKα inhibitor, ritanserin, including interactions at the atypical C1 domain distinct from the ATP binding region. Unexpectedly, deconstruction of ritanserin yielded a fragment compound that blocks DGKα activity through a conserved binding mode and enhanced selectivity against the kinome. Collectively, our studies illustrate the power of chemical proteomics to profile protein-small molecule interactions of lipid kinases for fragment-based lead discovery.
KW - activity-based protein profiling
KW - cancer immunotherapy
KW - chemical proteomics
KW - diacylglycerol kinase
KW - diacylglycerol signaling
KW - fragment-based drug discovery
KW - ligand deconstruction
KW - lipid kinase inhibitors
KW - lipid phosphorylation
KW - phosphatidic acid signaling
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U2 - 10.1016/j.chembiol.2017.06.007
DO - 10.1016/j.chembiol.2017.06.007
M3 - Article
C2 - 28712745
AN - SCOPUS:85023624992
SN - 2451-9456
VL - 24
SP - 870-880.e5
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 7
ER -
