Peptide array based screening reveals a large number of proteins interacting with the ankyrin repeat domain of the zDHHC17 S-acyltransferase

Lemonidis, K., MacLeod, R., Baillie, G. S. and Chamberlain, L. H. (2017) Peptide array based screening reveals a large number of proteins interacting with the ankyrin repeat domain of the zDHHC17 S-acyltransferase. Journal of Biological Chemistry, 292(42), pp. 17190-17202. (doi: 10.1074/jbc.M117.799650)

Abstract

zDHHC S-acyl-transferases are enzymes catalyzing protein S-acylation, a common post-translational modification on proteins frequently affecting their membrane targeting and trafficking. The ankyrin-repeat (AR) domain of zDHHC17 (HIP14) and zDHHC13 (HIP14L) S-acyltransferases, which is involved in both substrate recruitment and S-acylation-independent functions, was recently shown to bind at least six proteins, by specific recognition of a consensus sequence in them. To further refine the rules governing binding to the AR of zDHHC17, we employed peptide arrays based on zDHHC AR-binding motif (zDABM) sequences of synaptosomal-associated protein 25 (SNAP25) and cysteine string protein alpha (CSPα). Quantitative comparisons of the binding preferences of 400 peptides allowed us to construct a position-specific scoring matrix (PSSM) for zDHHC17 AR binding, with which we predicted and subsequently validated many putative zDHHC17 interactors. We identified 95 human zDABM sequences with unexpected versatility in amino-acid usage; these sequences were distributed among 90 proteins, of which 62 have not been previously implicated in zDHHC17/13 binding. These zDABM-containing proteins included all family members of the SNAP25, sprouty, cornifelin, ankyrin, and SLAIN-motif containing families; seven endogenous Gag polyproteins sharing the same binding sequence; and several proteins involved in cytoskeletal organization, cell communication, and regulation of signaling. A dozen of the zDABM-containing proteins had more than one zDABM sequence, while isoform-specific binding to the AR of zDHHC17 was identified for the Ena/VASP-like protein. The large number of zDABM sequences within the human proteome suggests that zDHHC17 may be an interaction hub regulating many cellular processes.

• Item Type: Articles
• Additional Information: This work was supported by grants from the BBSRC to LHC (BB/L022087/1 and BB/J006432/1).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Baillie, Professor George and MacLeod, Miss Ruth
• Authors: Lemonidis, K., MacLeod, R., Baillie, G. S., and Chamberlain, L. H.
• College/School: College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health
• Journal Name: Journal of Biological Chemistry
• Publisher: American Society for Biochemistry and Molecular Biology
• ISSN: 0021-9258
• ISSN (Online): 1083-351X
• Published Online: 07 September 2017
• Copyright Holders: Copyright © 2017 The Authors
• First Published: First published in Journal of Biological Chemistry 292(42):17190-17202
• Publisher Policy: Reproduced under a Creative Commons License