Molecular basis of fatty acid selectivity in the zDHHC family of S-acyltransferases revealed by click chemistry

Jennifer Greaves, Kevin R. Munro, Stuart C. Davidson, Matthieu Riviere, Justyna Wojno, Terry K. Smith, Nicholas C.O. Tomkinson, Luke H. Chamberlain

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

S-Acylation is a major post-translational modification, catalysed by the zDHHC enzyme family. S-acylated proteins can be modified by different fatty acids; however, very little is known about how zDHHC enzymes contribute to acyl chain heterogeneity. Here, we employed fatty acid azide/alkyne labelling of mammalian cells, showing their transformation into acyl-CoAs and subsequent click chemistry-based detection, to demonstrate that zDHHC enzymes have marked differences in their fatty acid selectivity. This was apparent even for highly related enzymes such as zDHHC3 and zDHHC7, which displayed a marked difference in ability to use C18:0 acyl CoA as a substrate. Furthermore, we identified Isoleucine-182 in the third transmembrane domain of zDHHC3 as a key determinant limiting the use of longer chain acyl-CoAs by this enzyme. This is the first study to uncover differences in the fatty acid selectivity profiles of cellular zDHHC enzymes and to map molecular determinants governing this selectivity
LanguageEnglish
Number of pages10
JournalProceedings of the National Academy of Sciences
Volume114
Issue number8
Early online date6 Feb 2017
DOIs
Publication statusE-pub ahead of print - 6 Feb 2017

Fingerprint

Click Chemistry
Acyltransferases
Fatty Acids
Enzymes
Acyl Coenzyme A
Alkynes
Acylation
Azides
Isoleucine
Protein S
Post Translational Protein Processing

Keywords

  • S-acylation
  • palmitoylation
  • zDHHC
  • click chemistry
  • fatty acid
  • acyl-CoA

Cite this

Greaves, Jennifer ; Munro, Kevin R. ; Davidson, Stuart C. ; Riviere, Matthieu ; Wojno, Justyna ; Smith, Terry K. ; Tomkinson, Nicholas C.O. ; Chamberlain, Luke H. / Molecular basis of fatty acid selectivity in the zDHHC family of S-acyltransferases revealed by click chemistry. In: Proceedings of the National Academy of Sciences . 2017 ; Vol. 114, No. 8.
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Molecular basis of fatty acid selectivity in the zDHHC family of S-acyltransferases revealed by click chemistry. / Greaves, Jennifer; Munro, Kevin R.; Davidson, Stuart C.; Riviere, Matthieu; Wojno, Justyna; Smith, Terry K.; Tomkinson, Nicholas C.O.; Chamberlain, Luke H.

In: Proceedings of the National Academy of Sciences , Vol. 114, No. 8, 06.02.2017.

Research output: Contribution to journalArticle

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AU - Smith, Terry K.

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