PXD017321

PXD017321 is an original dataset announced via ProteomeXchange.

Title	Proteomics Using Proteases Alternative to Trypsin Benefits from Sequential Digestion with Trypsin
Description	High specificity and ease of use make trypsin the most used enzyme in proteomics. Proteases with complementary cleavage specificity to trypsin have been applied to obtain additional data. However, use of proteases with broad specificity proved especially challenging. In this work, we analyzed the characteristics of five protease alternatives to trypsin for protein 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, the addition of trypsin to a proteinase K digest increased the number of identified proteins by 524 %. Also, AspN (82 %) and GluC (74 %) protein identifications largely improved following the additional digestion with trypsin. In general, protein identifications improved most over the use of the single protease when the enzymes followed on an initial digestion with trypsin. In the most extreme case, the sequential digest with trypsin and AspN yielded even higher number of protein identifications than digesting with trypsin alone.
HostingRepository	PRIDE
AnnounceDate	2021-06-07
AnnouncementXML	Submission_2021-06-07_05:10:36.842.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Therese Dau
SpeciesList	scientific name: Schizosaccharomyces pombe OY26; NCBI TaxID: 649908;
ModificationList	carbamoylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2020-01-29 05:33:20	ID requested
⏵ 1	2021-06-07 05:10:37	announced

Dau T, Bartolomucci G, Rappsilber J, Proteomics Using Protease Alternatives to Trypsin Benefits from Sequential Digestion with Trypsin. Anal Chem, 92(14):9523-9527(2020) [pubmed]

submitter keyword: sequential digest, alternative proteases

Juri Rappsilber
contact affiliation	Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355, Berlin, Germany Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3BF, Scotland, UK
contact email	juri.rappsilber@tu-berlin.de
lab head
Therese Dau
contact affiliation	University of Edinburgh
contact email	therese.dau@ed.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD017321
     1. Label: PRIDE project
     2. Name: Proteomics Using Proteases Alternative to Trypsin Benefits from Sequential Digestion with Trypsin