RapiGest precipitation depends on peptide concentration.
Peter R MosenRobert HardtDominic WinterPublished in: Proteomics (2021)
The mass spectrometry-compatible surfactant RapiGest promotes the enzymatic digestion of proteins by facilitating their unfolding while retaining enzymatic activity. RapiGest consists of a hydrophilic head and a hydrophobic tail, which can be separated by acid hydrolysis. This allows for removal of RapiGest prior to mass spectrometric analysis via precipitation and solid phase extraction. During in-solution digestion experiments with RapiGest, we noticed a high variability in the formation of precipitates after acid hydrolysis, implying that RapiGest precipitation is sample-dependent. We show that RapiGest hydrolyses efficiently under acidic conditions and that differences in precipitation are solely due to protein/peptide concentration. Furthermore, we demonstrate that RapiGest precipitation can be triggered by the addition of intact proteins, providing a strategy for its efficient removal from highly diluted samples. Data are available via ProteomeXchange with identifier PXD025982.
Keyphrases
- solid phase extraction
- liquid chromatography
- mass spectrometry
- high performance liquid chromatography
- anaerobic digestion
- liquid chromatography tandem mass spectrometry
- gas chromatography
- hydrogen peroxide
- molecularly imprinted
- simultaneous determination
- ionic liquid
- high resolution mass spectrometry
- gas chromatography mass spectrometry
- machine learning
- amino acid
- big data
- small molecule
- nitric oxide
- artificial intelligence
- capillary electrophoresis
- protein protein
- optical coherence tomography