Proteomics

Dataset Information

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Methylated trypsin - Enhanced trypsin on a budget: Stabilization, purification and high-temperature application of inexpensive commercial trypsin for proteomics applications


ABSTRACT: HCP analysis of the various trypsins was performed using the X! Tandem search engine (Alanine) (http://www.thegpm.org). All data was searched against the downloaded UniProt Sus scrofa proteome (40.706 sequences, March 10, 2019) and the most recent cRAP list (http://www.thegpm.org). Data were search as: enzyme: semi-tryptic, parent ion error: 10 ppm, fragment error: 0.1 Da, max e-value: 0.01, fixed modifications: carbamidomethylation (C), partial modifications: oxidation (M), methylation (K), dimethylation (K). Each triplicate set of result files were then analyzed through PeptideShaker 1.16.38 [26] and reported at 1% FDR.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

SUBMITTER: Sigurd Josef Frederiksen  

LAB HEAD: Peter Højrup

PROVIDER: PXD013458 | Pride | 2019-06-06

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
Pig_uniprot_crap.fasta Fasta
Pig_uniprot_crap_concatenated_target_decoy.fasta Fasta
QEHF3_06611_SJF_0.mgf Mgf
QEHF3_06611_SJF_0.raw Raw
QEHF3_06611_SJF_0.t.xml Xml
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Publications

Enhanced trypsin on a budget: Stabilization, purification and high-temperature application of inexpensive commercial trypsin for proteomics applications.

Heissel Søren S   Frederiksen Sigurd J SJ   Bunkenborg Jakob J   Højrup Peter P  

PloS one 20190627 6


Trypsin is by far the most commonly used protease in proteomics. Even though the amount of protease used in each experiment is very small, digestion of large amounts of protein prior to enrichment can be rather costly. The price of commercial trypsin is highly dependent on the quality of the enzyme, which is determined by its purity, activity, and chemical modifications. In this study we evaluated several strategies for improving the quality of crude trypsin by reductive methylation and affinity  ...[more]

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