Bivalent recognition of fatty acyl-CoA by a human integral membrane palmitoyltransferase.
DHHC acyltransferase
fatty acyl-CoA
integral membrane enzyme
membrane protein structure
protein S-acylation
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
15 02 2022
15 02 2022
Historique:
accepted:
21
12
2021
entrez:
10
2
2022
pubmed:
11
2
2022
medline:
11
3
2022
Statut:
ppublish
Résumé
S-acylation, also known as palmitoylation, is the most abundant form of protein lipidation in humans. This reversible posttranslational modification, which targets thousands of proteins, is catalyzed by 23 members of the DHHC family of integral membrane enzymes. DHHC enzymes use fatty acyl-CoA as the ubiquitous fatty acyl donor and become autoacylated at a catalytic cysteine; this intermediate subsequently transfers the fatty acyl group to a cysteine in the target protein. Protein S-acylation intersects with almost all areas of human physiology, and several DHHC enzymes are considered as possible therapeutic targets against diseases such as cancer. These efforts would greatly benefit from a detailed understanding of the molecular basis for this crucial enzymatic reaction. Here, we combine X-ray crystallography with all-atom molecular dynamics simulations to elucidate the structure of the precatalytic complex of human DHHC20 in complex with palmitoyl CoA. The resulting structure reveals that the fatty acyl chain inserts into a hydrophobic pocket within the transmembrane spanning region of the protein, whereas the CoA headgroup is recognized by the cytosolic domain through polar and ionic interactions. Biochemical experiments corroborate the predictions from our structural model. We show, using both computational and experimental analyses, that palmitoyl CoA acts as a bivalent ligand where the interaction of the DHHC enzyme with both the fatty acyl chain and the CoA headgroup is important for catalytic chemistry to proceed. This bivalency explains how, in the presence of high concentrations of free CoA under physiological conditions, DHHC enzymes can efficiently use palmitoyl CoA as a substrate for autoacylation.
Identifiants
pubmed: 35140179
pii: 2022050119
doi: 10.1073/pnas.2022050119
pmc: PMC8851515
pii:
doi:
Substances chimiques
Acyl Coenzyme A
0
Acyltransferases
EC 2.3.-
ZDHHC20 protein, human
EC 2.3.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Intramural NIH HHS
ID : ZIA HD008928
Pays : United States
Organisme : NIH HHS
ID : S10 OD012289
Pays : United States
Organisme : NIH HHS
ID : S10 OD021527
Pays : United States
Organisme : NIGMS NIH HHS
ID : P30 GM138396
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA DK036139
Pays : United States
Organisme : NIGMS NIH HHS
ID : P30 GM124165
Pays : United States
Informations de copyright
Copyright © 2022 the Author(s). Published by PNAS.
Déclaration de conflit d'intérêts
The authors declare no competing interest.
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