TY - JOUR
T1 - High-Throughput Enzyme Assay for Screening Inhibitors of the ZDHHC3/7/20 Acyltransferases
AU - Hong, Jun Young
AU - Malgapo, Martin Ian P.
AU - Liu, Yinong
AU - Yang, Min
AU - Zhu, Chengliang
AU - Zhang, Xiaoyu
AU - Tolbert, Patricia
AU - Linder, Maurine E.
AU - Lin, Hening
N1 - Funding Information:
This work is supported by an R01GM121540, a research grant from the National Institutes of Health. This work had use of the Cornell University NMR facility, which is supported, in part, by the NSF through MRI award CHE-1531632.
Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society.
PY - 2021/8/20
Y1 - 2021/8/20
N2 - As enzymes that mediate the attachment of long-chain fatty acids to cysteine residues, ZDHHC proteins have been reported to be promising therapeutic targets for treating cancer and autoimmune diseases. Yet, due to the lack of potent selective inhibitors, scrutiny of the biological functions of ZDHHCs has been limited. The main hindrance for developing ZDHHC inhibitors is the lack of a facile high-throughput assay. Here, we developed a ZDHHC3/7/20 high-throughput assay based on the acylation-coupled lipophilic induction of polarization (Acyl-cLIP) method and screened several potential ZDHHC inhibitors. Furthermore, we demonstrated that in vitro results from the Acyl-cLIP assay are supported by the results from cell-based assays. We envision that this new ZDHHC3/7/20 Acyl-cLIP assay will accelerate the high-throughput screening of large compound libraries for improved ZDHHC inhibitors and provide therapeutic benefits for cancer and autoimmune diseases.
AB - As enzymes that mediate the attachment of long-chain fatty acids to cysteine residues, ZDHHC proteins have been reported to be promising therapeutic targets for treating cancer and autoimmune diseases. Yet, due to the lack of potent selective inhibitors, scrutiny of the biological functions of ZDHHCs has been limited. The main hindrance for developing ZDHHC inhibitors is the lack of a facile high-throughput assay. Here, we developed a ZDHHC3/7/20 high-throughput assay based on the acylation-coupled lipophilic induction of polarization (Acyl-cLIP) method and screened several potential ZDHHC inhibitors. Furthermore, we demonstrated that in vitro results from the Acyl-cLIP assay are supported by the results from cell-based assays. We envision that this new ZDHHC3/7/20 Acyl-cLIP assay will accelerate the high-throughput screening of large compound libraries for improved ZDHHC inhibitors and provide therapeutic benefits for cancer and autoimmune diseases.
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U2 - 10.1021/acschembio.1c00258
DO - 10.1021/acschembio.1c00258
M3 - Article
C2 - 34374518
AN - SCOPUS:85113950319
SN - 1554-8929
VL - 16
SP - 1318
EP - 1324
JO - ACS chemical biology
JF - ACS chemical biology
IS - 8
ER -
