Detection of Protein S-Acylation using Acyl-Resin Assisted Capture
| Autor: | Savannah J. West, Askar M. Akimzhanov, Ritika Tewari, Bieerkehazi Shayahati |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification General Immunology and Microbiology Acylation General Chemical Engineering General Neuroscience Fatty acid S-acylation Biological activity Thioester Article General Biochemistry Genetics and Molecular Biology Protein S Sepharose 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Palmitoylation chemistry Biochemistry lipids (amino acids peptides and proteins) 030217 neurology & neurosurgery Cysteine |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| DOI: | 10.3791/61016-v |
| Popis: | Protein S-acylation, also referred to as S-palmitoylation, is a reversible post-translational modification of cysteine residues with long-chain fatty acids via a labile thioester bond. S-acylation, which is emerging as a widespread regulatory mechanism, can modulate almost all aspects of the biological activity of proteins, from complex formation to protein trafficking and protein stability. The recent progress in understanding of the biological function of protein S-acylation was achieved largely due to the development of novel biochemical tools allowing robust and sensitive detection of protein S-acylation in a variety of biological samples. Here, we describe acyl resin-assisted capture (Acyl-RAC), a recently developed method based on selective capture of endogenously S-acylated proteins by thiol-reactive Sepharose beads. Compared to existing approaches, Acyl-RAC requires fewer steps and can yield more reliable results when coupled with mass spectrometry for identification of novel S-acylation targets. A major limitation in this technique is the lack of ability to discriminate between fatty acid species attached to cysteines via the same thioester bond. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|