Quantitative analysis of protein lipidation and acyl-CoAs reveals substrate preferences of the S -acylation machinery
Busquets-Hernández, Carla and Ribó, Silvia and Gratacós-Batlle, Esther and Carbajo, Daniel and Tsiotsia, Alexandra and Blanco-Canosa, Juan B. and Chamberlain, Luke H. and Triola, Gemma (2024) Quantitative analysis of protein lipidation and acyl-CoAs reveals substrate preferences of the S -acylation machinery. Chemical Science, 15 (32). pp. 12845-12855. ISSN 2041-6539 (https://doi.org/10.1039/d4sc02235a)
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Abstract
Protein palmitoylation or S-acylation has emerged as a key regulator of cellular processes. Increasing evidence shows that this modification is not restricted to palmitate but it can include additional fatty acids, raising the possibility that differential S-acylation contributes to the fine-tuning of protein activity. However, methods to profile the acyl moieties attached to proteins are scarce. Herein, we report a method for the identification and quantification of lipids bound to proteins that relies on hydroxylamine treatment and mass spectrometry analysis of fatty acid hydroxamates. This method has enabled unprecedented and extensive profiling of the S-acylome in different cell lines and tissues and has shed light on the substrate specificity of some S-acylating enzymes. Moreover, we could extend it to quantify also the acyl-CoAs, which are thioesters formed between a fatty acid and a coenzyme A, overcoming many of the previously described challenges for the detection of such species. Importantly, the simultaneous analysis of the lipid fraction and the proteome allowed us to establish, for the first time, a direct correlation between the endogenous levels of acyl-CoAs and the S-acylation profile of its proteome.
ORCID iDs
Busquets-Hernández, Carla, Ribó, Silvia, Gratacós-Batlle, Esther, Carbajo, Daniel, Tsiotsia, Alexandra, Blanco-Canosa, Juan B., Chamberlain, Luke H. ORCID: https://orcid.org/0000-0002-8701-4995 and Triola, Gemma;-
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Item type: Article ID code: 89936 Dates: DateEvent28 August 2024Published9 July 2024Published Online8 July 2024Accepted4 April 2024SubmittedSubjects: Science > Chemistry Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences Depositing user: Pure Administrator Date deposited: 16 Jul 2024 14:42 Last modified: 01 Oct 2024 09:44 URI: https://strathprints.strath.ac.uk/id/eprint/89936