Bai, M. et al. (2024) Targeted degradation of zDHHC-PATs decreases substrate S -palmitoylation. PLoS ONE, 19(3), e0299665. (doi: 10.1371/journal.pone.0299665) (PMID:38512906) (PMCID:PMC10956751)
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Abstract
Reversible S-palmitoylation of protein cysteines, catalysed by a family of integral membrane zDHHC-motif containing palmitoyl acyl transferases (zDHHC-PATs), controls the localisation, activity, and interactions of numerous integral and peripheral membrane proteins. There are compelling reasons to want to inhibit the activity of individual zDHHC-PATs in both the laboratory and the clinic, but the specificity of existing tools is poor. Given the extensive conservation of the zDHHC-PAT active site, development of isoform-specific competitive inhibitors is highly challenging. We therefore hypothesised that proteolysis-targeting chimaeras (PROTACs) may offer greater specificity to target this class of enzymes. In proof-of-principle experiments we engineered cell lines expressing tetracycline-inducible Halo-tagged zDHHC5 or zDHHC20, and evaluated the impact of Halo-PROTACs on zDHHC-PAT expression and substrate palmitoylation. In HEK-derived FT-293 cells, Halo-zDHHC5 degradation significantly decreased palmitoylation of its substrate phospholemman, and Halo-zDHHC20 degradation significantly diminished palmitoylation of its substrate IFITM3, but not of the SARS-CoV-2 spike protein. In contrast, in a second kidney derived cell line, Vero E6, Halo-zDHHC20 degradation did not alter palmitoylation of either IFITM3 or SARS-CoV-2 spike. We conclude from these experiments that PROTAC-mediated targeting of zDHHC-PATs to decrease substrate palmitoylation is feasible. However, given the well-established degeneracy in the zDHHC-PAT family, in some settings the activity of non-targeted zDHHC-PATs may substitute and preserve substrate palmitoylation.
Item Type: | Articles |
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Additional Information: | Funding: We acknowledge support from the following funders: British Heart Foundation, RG/17/15/33106 Professor William Fuller Medical Research Scotland, Daphne Jackson Fellowship Simon Swingler |
Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Howie, Dr Jacqueline and Swingler, Dr Simon and Fuller, Professor Will and GAO, Xing and Kuo, Dr Chien-Wen and Wilson, Professor Sam and France, Dr David and Memarzadeh, Ms Sarah and Gallen, Emily and Brown, Miss Elaine |
Creator Roles: | France, D.Conceptualization, Funding acquisition, Supervision, Writing – original draft, Writing – review and editing Fuller, W.Conceptualization, Funding acquisition, Supervision, Writing – original draft, Writing – review and editing Gallen, E.Investigation Memarzadeh, S.Investigation, Writing – original draft, Writing – review and editing Howie, J.Investigation GAO, X.Investigation Kuo, C.-W.Investigation Brown, E.Methodology Swingler, S.Methodology Wilson, S.Supervision, Writing – review and editing |
Authors: | Bai, M., Gallen, E., Memarzadeh, S., Howie, J., GAO, X., Kuo, C.-W. S., Brown, E., Swingler, S., Wilson, S. J., Shattock, M. J., France, D. J., and Fuller, W. |
College/School: | College of Medical Veterinary and Life Sciences College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Science and Engineering > School of Chemistry |
Journal Name: | PLoS ONE |
Publisher: | Public Library of Science |
ISSN: | 1932-6203 |
ISSN (Online): | 1932-6203 |
Copyright Holders: | Copyright: © 2024 Bai et al. |
First Published: | First published in PLoS ONE 19(3): e0299665 |
Publisher Policy: | Reproduced under a Creative Commons licence |
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