Projects per year
Abstract
Trypsin is the most used enzyme in proteomics. Nevertheless, proteases with complementary cleavage specificity have been applied in special circumstances. In this work, we analyzed the characteristics of five protease alternatives to trypsin for pro-tein identification and sequence coverage when applied to S. pombe whole cell lysates. The specificity of the protease heavily impacted on the number of proteins identified. Proteases with higher specificity let to the identification of more proteins than proteases with lower specificity. However, AspN, GluC, chymotrypsin and proteinase K largely benefited from being paired with trypsin in sequential digestion, as had been shown by us for elastase before. In the most extreme case, pre-digesting with trypsin improves the number of identified proteins for proteinase K by 731 %. Trypsin pre-digestion also improved the protein identifications of other proteases, AspN (+62 %), GluC (+80 %) and chymotrypsin (+21 %). Interestingly, the sequential digest with trypsin and AspN yielded even higher number of protein identifications than digesting with trypsin alone.
Original language | English |
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Journal | Analytical Chemistry |
Early online date | 6 Jul 2020 |
DOIs | |
Publication status | E-pub ahead of print - 6 Jul 2020 |
Keywords / Materials (for Non-textual outputs)
- sequential digest
- alternative proteases
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Dive into the research topics of 'Proteomics using protease alternatives to trypsin benefits from sequential digestion with trypsin'. Together they form a unique fingerprint.Projects
- 4 Finished
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`Core Funding for the Wellcome Trust Centre for Cell Biology¿, Research Enrichment, Public Engagement
1/12/18 → 1/06/22
Project: Research
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Datasets
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Sequential digestion with Trypsin and Elastase in cross-linking mass spectrometry
Dau, T. (Creator), Gupta, K. (Creator), Berger, I. (Creator) & Rappsilber, J. (Creator), PRIDE database hosted by European Bioinformatics Institute, EBI, 7 Mar 2019
Dataset