A Palmitoyl Transferase Chemical-genetic System to Map ZDHHC-specific S-acylation

Overview

Journal Nat Biotechnol

Specialty Biotechnology

Date 2024 Jan 8

PMID 38191663

Authors

Cory A Ocasio

Marc P Baggelaar

James Sipthorp

Ana Losada de la Lastra

Manuel Tavares

Jana Volaric

Christelle Soudy

Elisabeth M Storck

Jack W Houghton

Susana A Palma-Duran

James I MacRae

Goran Tomic

Lotte Carr

Julian Downward

Ulrike S Eggert

Edward W Tate

Affiliations

Soon will be listed here.

Abstract

The 23 human zinc finger Asp-His-His-Cys motif-containing (ZDHHC) S-acyltransferases catalyze long-chain S-acylation at cysteine residues across an extensive network of hundreds of proteins important for normal physiology or dysregulated in disease. Here we present a technology to directly map the protein substrates of a specific ZDHHC at the whole-proteome level, in intact cells. Structure-guided engineering of paired ZDHHC 'hole' mutants and 'bumped' chemically tagged fatty acid probes enabled probe transfer to specific protein substrates with excellent selectivity over wild-type ZDHHCs. Chemical-genetic systems were exemplified for five human ZDHHCs (3, 7, 11, 15 and 20) and applied to generate de novo ZDHHC substrate profiles, identifying >300 substrates and S-acylation sites for new functionally diverse proteins across multiple cell lines. We expect that this platform will elucidate S-acylation biology for a wide range of models and organisms.

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