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. 2021 Oct;21(20):e2100129.
doi: 10.1002/pmic.202100129. Epub 2021 Sep 5.

RapiGest precipitation depends on peptide concentration

Peter R Mosen  1 , Robert Hardt  1 , Dominic Winter  1
Affiliations

RapiGest precipitation depends on peptide concentration

Peter R Mosen et al. Proteomics. 2021 Oct.

Abstract

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.

Keywords: RapiGest; in solution digestion; precipitation; proteomics; surfactant.

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References

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