Characterization of hedgehog acyltransferase inhibitors identifies a small molecule probe for hedgehog signaling by cancer cells
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
The Sonic Hedgehog (Shh) signaling pathway plays a critical role during embryonic development and cancer progression. N-terminal palmitoylation of Shh by Hedgehog acyltransferase (Hhat) is essential for efficient signaling, raising interest in Hhat as a novel drug target. A recently identified series of dihydrothienopyridines has been proposed to function via this mode of action; however, the lead compound in this series (RUSKI-43) was subsequently shown to possess cytotoxic activity unrelated to canonical Shh signaling. To identify a selective chemical probe for cellular studies, we profiled three RUSKI compounds in orthogonal cell-based assays. We found that RUSKI-43 exhibits off-target cytotoxicity, masking its effect on Hhat-dependent signaling, hence results obtained with this compound in cells should be treated with caution. In contrast, RUSKI-201 showed no off-target cytotoxicity, and quantitative whole-proteome palmitoylation profiling with a bioorthogonal alkyne-palmitate reporter demonstrated specific inhibition of Hhat in cells. RUSKI-201 is the first selective Hhat chemical probe in cells and should be used in future studies of Hhat catalytic function.
Date Issued
2016-10-25
Date Acceptance
2016-10-13
Citation
ACS Chemical Biology, 2016, 11 (12), pp.3256-3262
ISSN
1554-8937
Publisher
American Chemical Society
Start Page
3256
End Page
3262
Journal / Book Title
ACS Chemical Biology
Volume
11
Issue
12
Copyright Statement
© 2016 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Sponsor
Cancer Research UK
Commission of the European Communities
Grant Number
C6433/A16402
PIEF-GA-2013-623648
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
PANCREATIC DUCTAL ADENOCARCINOMA
SONIC-HEDGEHOG
CHEMICAL PROBES
PROTEIN LIPIDATION
TARGET VALIDATION
CLICK CHEMISTRY
PALMITOYLATION
RANGE
CHOLESTERYLATION
MODULATION
Acyltransferases
Animals
Cell Line, Tumor
Enzyme Inhibitors
HEK293 Cells
Hedgehog Proteins
Humans
Lipoylation
Mice
NIH 3T3 Cells
Neoplasms
Signal Transduction
Organic Chemistry
03 Chemical Sciences
06 Biological Sciences
Publication Status
Published