AI Article Synopsis
- Using multiple proteases in mass spectrometry-based proteomics allows for the identification of more peptides and proteins compared to using a single protease.
- Trypsin and Tryp-N target different sites on the same amino acids (C-terminal and N-terminal respectively), enhancing the overall detection of proteins.
- The study demonstrates that combining these proteases with S-Trap columns results in unique protein profiles from plasma and cell lysates, suggesting this dual approach yields better proteome coverage.
Article Abstract
Mass spectrometry-based proteomics combining more than one protease in parallel facilitates the identification of more peptides and proteins than when a single protease is used. Trypsin cleaves proteins C-terminally to arginine and lysine, while its mirroring protease Tryp-N cleaves N-terminally to the same amino acids. Here, we combine trypsin and Tryp-N with the commercially available S-Trap columns, which purify protein samples and catalyze digestion. Comparison of trypsin or Tryp-N coupled with S-Trap columns demonstrates plasma and cell lysate proteins unique to one protease. We thus suggest the use of both proteases in a complementary manner to obtain deeper proteome coverage.
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| http://dx.doi.org/10.1007/978-1-0716-3457-8_1 | DOI Listing |
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