Generating cysteine-trypsin cleavage sites with 2-chloroacetamidine capping.
Samuel OforiHeta S DesaiFlowreen ShikwanaLisa M BoatnerEmil R Dominguez IiiJosé O CastellónKeriann M BackusPublished in: Chemical communications (Cambridge, England) (2024)
An electrophilic arginine mimetic, 2-chloroacetamidine (CAM), was deployed to enable trypsin-mediated proteolysis at cysteine residues and to enhance mass spectrometry-based proteomic detection of cysteine-containing peptides. Illustrating the value of the CAM-capping strategy, proteogenomic analysis using a two-stage false discovery rate (FDR) search revealed >50% enhanced coverage of missense variants, when compared to established workflows.
Keyphrases
- fluorescent probe
- mass spectrometry
- living cells
- label free
- small molecule
- nitric oxide
- liquid chromatography
- copy number
- high throughput
- intellectual disability
- amino acid
- healthcare
- high performance liquid chromatography
- gene expression
- loop mediated isothermal amplification
- dna binding
- gas chromatography
- ms ms
- dna methylation
- autism spectrum disorder
- tandem mass spectrometry