PXD015612-1

PXD015612 is an original dataset announced via ProteomeXchange.

Title	Exopeptidase assisted N- and C-terminal proteome sequencing
Description	Due to mechanisms such as alternative translation starts or proteolytic processing, in-vivo proteoforms do not necessarily correspond to those encoded in the genome. Therefore, knowledge of protein termini is an indispensable prerequisite to understand protein function. Sequencing of protein N- and C-termini has been limited to single purified protein species, while the proteome-wide identification of N- and C-termini relies on the generation of single, terminal proteotypic peptides followed by chemical enrichment or depletion strategies to facilitate their detection via mass spectrometry (MS). To overcome limitations in such approaches, we present a novel concept that readily enables unbiased ladder sequencing of protein N- and C-termini. The approach is based on the combination of exopeptidase digestions and two-dimensional separation by strong cation exchange chromatography and reversed-phase liquid chromatography coupled online to tandem-MS. We demonstrate the potential of this methodology by analyzing the N- and C-terminome of S. cerevisiae, identifying 2,189 N-termini, and 1,562 C-termini. In conclusion, the presented method largely expands the proteomics toolbox enabling N- and C-terminal sequential characterization of entire proteomes.
HostingRepository	PRIDE
AnnounceDate	2025-10-16
AnnouncementXML	Submission_2025-10-16_01:41:20.752.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andreas Tholey
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: NEWT:4932;
ModificationList	(18)O label at both C-terminal oxygens; acetylated residue; dimethylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-09-27 00:21:38	ID requested
⏵ 1	2025-10-16 01:41:21	announced

Helbig AO, Tholey A, Exopeptidase Assisted N- and C-Terminal Proteome Sequencing. Anal Chem, 92(7):5023-5032(2020) [pubmed]

10.1021/acs.analchem.9b05288;

curator keyword: Biological

submitter keyword: Protein sequencing, Proteomics, Terminomics

Prof. Dr. Andreas Tholey
contact affiliation	Systematische Proteomforschung & Bioanalytik Institut für Experimentelle Medizin (IEM) Christian-Albrechts-Universität zu Kiel Niemannsweg 11 24105 Kiel, Germany
contact email	a.tholey@iem.uni-kiel.de
lab head
Andreas Tholey
contact affiliation	Systematic Proteome Research & Bioanalytics, University of Kiel
contact email	a.tholey@iem.uni-kiel.de
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/2025/10/PXD015612

PRIDE project URI

• PRIDE
  1. PXD015612
     1. Label: PRIDE project
     2. Name: Exopeptidase assisted N- and C-terminal proteome sequencing