PXD021191

PXD021191 is an original dataset announced via ProteomeXchange.

Title	ProAlanase is an effective alternative to trypsin for proteomics applications and disulfide bond mapping_part2
Description	Trypsin is the protease of choice in bottom-up proteomics. However, its application can be limited by the amino acid composition of target proteins and the pH of digestion solution. In this study we characterized ProAlanase, a protease from the fungus Aspergillus niger that cleaves primarily on the C-terminal side of proline and alanine residues. ProAlanase achieves high proteolytic activity and specificity when digestions are carried out at acidic pH (1.5) for relatively short (2 hr) time periods. To elucidate the potential of ProAlanase in proteomic applications, we conducted a series of investigations comprising digestion of proline-rich proteins, PTM analysis, de novo protein sequencing and disulfide bond mapping. The results demonstrated that digestion with ProAlanase improves protein sequence coverage and phosphosite localization for the proline-rich protein Notch3 intracellular domain and improves non-collagenous bone protein identification. Notably, cleavage also occurs at the C-terminus of hydroxyproline, facilitating efficient digestion of bone collagen. Finally, we demonstrate that ProAlanase is efficient in disulfide bond mapping, showing high coverage of disulfide-containing regions in monoclonal antibodies, as well as achieving nearly complete database-independent sequence reconstruction of endogenous protein by de novo sequencing.
HostingRepository	PRIDE
AnnounceDate	2020-10-19
AnnouncementXML	Submission_2020-10-19_00:21:30.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Diana Samodova
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monomethylated residue; phosphorylated residue; acetylated residue
Instrument	Orbitrap Exploris 480; Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2020-08-28 07:05:45	ID requested
⏵ 1	2020-10-19 00:21:31	announced

Samodova D, Hosfield CM, Cramer CN, Giuli MV, Cappellini E, Franciosa G, Rosenblatt MM, Kelstrup CD, Olsen JV, ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping. Mol Cell Proteomics, 19(12):2139-2157(2020) [pubmed]

submitter keyword: Proteomics, ProAlanase, sequence coverage, phosphorylation, proline-rich, low-pH, disulfide bonds, disulfide mapping, histone variants, histone PTMs, de novo sequencing

Jesper Velgaard Olsen
contact affiliation	Novo Nordisk Foundation Center for Protein Research
contact email	jesper.olsen@cpr.ku.dk
lab head
Diana Samodova
contact affiliation	University of Copenhagen, Novo Nordisk Foundation Center for Protein Research
contact email	diana.samodova@cpr.ku.dk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD021191
     1. Label: PRIDE project
     2. Name: ProAlanase is an effective alternative to trypsin for proteomics applications and disulfide bond mapping_part2