PXD030791

PXD030791 is an original dataset announced via ProteomeXchange.

Title	NAPT, an unbiased approach for sequential analysis of protein N- and C- terminome
Description	Robust analysis of both protein N- and C-termini can reveal novel N- and C-degrons that modify protein stability and shape the eukaryotic proteome. Herein, we designed a workflow, termed LysargiNase-assisted Analysis of Protein N- and C-Terminome (NAPT) by sequential and selective separating N- and C-termini on strong cation exchange chromatography (SCX). Taking advantage of N-terminal peptides’ reduced charge states at pH 2.7 and C-terminal peptides’ enhanced charge states at pH 8.5, an adequate shift of pH during SCX fractionation could easily lead to sequential elution of N-terminal, internal, and C-terminal peptides. Combining NAPT with multiple enzymatic digestion strategies, we identified 2458 human protein N-termini and 3056 human protein C-termini from Hela cell line. As N-terminal peptides generally bear two less positive charges than internal peptides at pH 2.7 after LysargiNase digestion, leaving one-positive-charge tolerance for histidine-containing N-terminal peptides, and C-terminal peptides were separated at pH 8.5 where histidine was neutrally charged, NAPT workflow showed minimal bias against histidine residues and excellent complementarity compared to its sister, the SAPT workflow, which adopted trypsin as protease. Taken together, 4285 protein N-termini and 4170 protein C-termini were identified by NAPT and SAPT, representing the largest human protein C-termini and the second largest human protein N-termini dataset so far, demonstrating their valuable potential in terminomic studies. Furthermore, we systematically analyzed sequences of identified protein termini and validated that their behavior obeyed several degron pathways.
HostingRepository	iProX
AnnounceDate	2022-04-11
AnnouncementXML	Submission_2023-12-21_22:34:51.750.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wenjuan Yuan
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-06 22:07:11	ID requested
1	2022-05-03 22:11:08	announced
⏵ 2	2023-12-21 22:34:52	announced	2023-12-22: Update information.

Wang Z, Yuan W, Zhang L, Zhang Y, Lu H, NAPT, an unbiased approach for sequential analysis of the protein N- and C-terminome. Chem Commun (Camb), 58(67):9397-9400(2022) [pubmed]

submitter keyword: hela cell, N- and C- terminome, LC-MS/MS

Haojie Lu
contact affiliation	Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University
contact email	luhaojie@fudan.edu.cn
lab head
Wenjuan Yuan
contact affiliation	Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai
contact email	13110220030@fudan.edu.cn
dataset submitter

iProX dataset URI