Thesis
S-Acylation of membrane proteins by the zDHHC family
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2026
- Thesis identifier
- T18168
- Person Identifier (Local)
- 202284275
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- S-Acylation is a reversible lipid modification crucial for regulating membrane protein trafficking, stability, and signalling, and catalysed by the zDHHC family of protein acyltransferases. While its importance is well-established, the substrate specificity of individual zDHHC enzymes and the rules governing S-acylation across different transmembrane protein classes remain incompletely understood. Building on recent discoveries that the endoplasmic reticulum enzyme zDHHC6 exhibits broad activity towards single-pass transmembrane proteins with membrane-proximal cysteines, this thesis investigates the S-acylation of multi-transmembrane proteins, including GPCRs and tetraspanins, focusing on their recognition by zDHHC6 in comparison to Golgilocalised enzymes zDHHC3 and zDHHC15. The first results chapter explores S-acylation of the GPCR CCR5, identifies new sites of S-acylation, shows that modification occurs after release from the endoplasmic reticulum, and demonstrates that zDHHC3 and zDHHC15, not zDHHC6, mediate this process. S-Acylation of CCR5 did not affect protein stability, whereas O-glycosylation did. The second results chapter analyses multi-transmembrane proteins, finding zDHHC6 activity against proteins with two and four transmembrane domains, but limited activity towards GPCRs, which are primarily substrates of zDHHC3 and zDHHC15. Mutational analysis rejected the idea that zDHHC6 specifically recognises membrane proximal cysteines in transmembrane proteins, revealing a more complex enzyme substrate interaction. S-Acylation of GPCRs on post-endoplasmic reticulum membranes was further supported by analysis of ER-trapped β2AR. Fatty acid attachment to transmembrane proteins followed the selectivity of their modifying enzymes, regardless of whether target cysteines were located in the cytosol or membrane. The third results chapter reports the novel S-acylation of a single-pass protein, TEMP, and its modification by zDHHC6. Overall, this work expands the catalogue of S-acylated transmembrane proteins and provides new insight into the determinants of zDHHC enzyme specificity, highlighting the importance of subcellular localisation, protein architecture, and lipid selectivity in regulating S-acylation.
- Advisor / supervisor
- Chamberlain, Luke
- Resource Type
- DOI
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