EVALUATION OF RAPIGEST EFFICACY FOR THE DIGESTION OF PROTEINS FROM CELL CULTURES AND HEART TISSUE

Abstract

Introduction: Rapigest is an acid-labile detergent used in proteomics for the improvement of protein digestion.

Materials and Method: To test the efficacy of Rapigest for proteomics analysis of different sample types we used protein extracts from S9 cell line and mouse heart tissue and performed protein isolation, digestion and mass spectrometry analysis.

Results: For the S9 cell line, there was no significant difference concerning the number of identifications (peptides, proteins) between Rapigest and No Rapigest samples, though slightly more peptides and proteins were identified in the Rapigest samples. For the mouse heart tissue samples, Rapigest use resulted in the identification of a higher number of proteins. Rapigest did not modify the protein profile with respect to the biological compartments covered by the identified proteins in S9 cell line samples, but produced a small increase in the representation of cytoplasm proteins and a small decrease in the representation of membrane proteins in the mouse heart tissue samples.

Discussions: Results are comparable to other studies that evaluated the efficacy of Rapigest for the analysis of tissue samples, recommending Rapigest for the improvement of protein digestion and implicitly identification, without the modification of the protein profile in the samples.

Conclusion: Rapigest may be successfully used for the improvement of protein identification from heart tissue samples using mass spectrometry.