PXD035682

PXD035682 is an original dataset announced via ProteomeXchange.

Title	Rapid trypsin digestion at elevated temperature with calcium for enhanced bottom-up proteome analysis
Description	Trypsin is frequently employed to cleave proteins ahead of mass spectrometry characterization. Traditionally, enzyme digestion involves overnight incubation of proteins at 37 °C, which is not only time consuming but can still yield in poor digestion efficiency. While raising the temperature should theoretically accelerate the digestion, this also destabilizes the enzyme and enhances trypsin degradation. We therefore questioned whether elevated temperature digestion is beneficial for improving tryptic digestion. Here we quantify protein digestion kinetics at elevated temperatures for calcium-stabilized trypsin and enforce the critical importance of calcium ions to preserve the enzyme. We quantitatively demonstrate that 1 hour at 47 °C provides a superior digest when compared to conventional (overnight, 37 °C) processing of the proteome. The impact of our enhanced digestion protocol is shown through bottom-up mass spectrometry analysis of a complex proteome mixture.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:41:23.156.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jessica Nickerson
SpeciesList	scientific name: Bacteria; NCBI TaxID: NCBITaxon:2; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-01 09:16:57	ID requested
1	2023-03-11 03:09:30	announced
⏵ 2	2023-11-14 08:41:26	announced	2023-11-14: Updated project metadata.

Nickerson JL, Doucette AA, Maximizing Cumulative Trypsin Activity with Calcium at Elevated Temperature for Enhanced Bottom-Up Proteome Analysis. Biology (Basel), 11(10):(2022) [pubmed]

submitter keyword: trypsin

calcium

bottom-up proteomics

mass spectrometry

accelerated digestion

digestion efficiency

quantitative proteomics

sample preparation

Alan Austin Doucette
contact affiliation	Department of Chemistry, Dalhousie University, Halifax, Canada
contact email	alan.doucette@dal.ca
lab head
Jessica Nickerson
contact affiliation	Dalhousie University
contact email	jessica.nickerson@dal.ca
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/03/PXD035682

PRIDE project URI

• PRIDE
  1. PXD035682
     1. Label: PRIDE project
     2. Name: Rapid trypsin digestion at elevated temperature with calcium for enhanced bottom-up proteome analysis