Analysis of Protein Cysteine Acylation Using a Modified Suspension Trap (Acyl-Trap).

ABSTRACT: Proteins undergo reversible S-acylation via a thioester linkage in vivo. S-palmitoylation, modification by C16:0 fatty acid, is a common S-acylation that mediates critical protein-membrane and protein-protein interactions. The most widely used S-acylation assays, including acyl-biotin exchange and acyl resin-assisted capture, utilize blocking of free Cys thiols, hydroxylamine-dependent cleavage of the thioester and subsequent labeling of nascent thiol. These assays generally require >500 micrograms of protein input material per sample and numerous reagent removal and washing steps, making them laborious and ill-suited for high throughput and low input applications. To overcome these limitations, we devised "Acyl-Trap", a suspension trap-based assay that utilizes

SUBMITTER: Forrester MT 

PROVIDER: S-EPMC10996552 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature