S-Acylation of Sprouty and SPRED proteins by the S-acyltransferase zDHHC17 involves a novel mode of enzyme-substrate interaction

Butler, L., Locatelli, C., Allagioti, D., Lousa, I., Lemonidis, K. , Tomkinson, N. C.O., Salaun, C. and Chamberlain, L. H. (2022) S-Acylation of Sprouty and SPRED proteins by the S-acyltransferase zDHHC17 involves a novel mode of enzyme-substrate interaction. Journal of Biological Chemistry, 299(1), 102754. (doi: 10.1016/j.jbc.2022.102754) (PMID:36442513) (PMCID:PMC9800311)

Abstract

S-Acylation is an essential post-translational modification, which is mediated by a family of twenty-three zDHHC enzymes in humans. Several thousand proteins are modified by S-acylation; however, we lack a detailed understanding of how enzyme-substrate recognition and specificity is achieved. Previous work showed that the ankyrin repeat domain of zDHHC17 (ANK17) recognizes a short linear motif, known as the zDHHC ANK binding motif (zDABM) in substrate protein SNAP25, as a mechanism of substrate recruitment prior to S-acylation. Here, we investigated the S-acylation of the Sprouty and SPRED family of proteins by zDHHC17. Interestingly, although Sprouty-2 (Spry2) contains a zDABM that interacts with ANK17, this mode of binding is dispensable for S-acylation, and indeed removal of the zDABM does not completely ablate binding to zDHHC17. Furthermore, the related SPRED3 protein interacts with and is efficiently S-acylated by zDHHC17 despite lacking a zDABM. We undertook mutational analysis of SPRED3 to better understand the basis of its zDABM-independent interaction with zDHHC17. This analysis found that the cysteine-rich SPR domain of SPRED3, which is the defining feature of all Sprouty and SPRED proteins, interacts with zDHHC17. Surprisingly, the interaction with SPRED3 was independent of ANK17. Our mutational analysis of Spry2 was consistent with the SPR domain of this protein containing a zDHHC17 binding site, and Srpy2 also showed detectable binding to a zDHHC17 mutant lacking the ANK domain. Thus, zDHHC17 can recognize its substrates through ANK domain and zDABM-dependent and –independent mechanisms, and some substrates display more than one mode of binding to this enzyme.

• Item Type: Articles
• Additional Information: This work was funded by grants from the BBSRC (BB/L022087/1) and the MRC (MR/R011842/1).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Lemonidis, Dr Kimon and Tomkinson, Dr Nicholas
• Authors: Butler, L., Locatelli, C., Allagioti, D., Lousa, I., Lemonidis, K., Tomkinson, N. C.O., Salaun, C., and Chamberlain, L. H.
• College/School: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences; College of Science and Engineering > School of Chemistry
• Journal Name: Journal of Biological Chemistry
• Publisher: American Society for Biochemistry and Molecular Biology, Inc.
• ISSN: 0021-9258
• ISSN (Online): 1083-351X
• Published Online: 25 November 2022
• Copyright Holders: Copyright © 2022 The Authors
• First Published: First published in Journal of Biological Chemistry 299(1):102754
• Publisher Policy: Reproduced under a Creative Commons License