PXD022988-1

PXD022988 is an original dataset announced via ProteomeXchange.

Title	Counterion Optimization Dramatically Improves Selectivity for Phosphopeptides and Glycopeptides in Electrostatic Repulsion-Hydrophilic Interaction Chromatography (ERLIC)
Description	A well-hydrated counterion can selectively and dramatically increase retention of a charged analyte in hydrophilic interaction chromatography (HILIC). The effect is enhanced if the column is charged, as in electrostatic repulsion-hydrophilic interaction chromatography (ERLIC). This combination was exploited in proteomics for the isolation of peptides with certain post-translational modifications (PTMs). The best salt additive examined was magnesium trifluoroacetate. The well-hydrated Mg+2 ion promoted retention of peptides with functional groups that retained negative charge at low pH, while the poorly-hydrated trifluoroacetate counterion tuned down the retention due to the basic residues. The result was an enhancement in selectivity between 6- to 66-fold. These conditions were applied to a tryptic digest of mouse cortex. Gradient elution produced fractions enriched in peptides with phosphate, mannose-6-phosphate, N- and O-linked glycans. The numbers of such peptides identified either equaled or exceeded the numbers afforded by the best alternative methods. This method is a productive and convenient way to isolate peptides simultaneously that contain a number of different PTMs, facilitating study of proteins with “crosstalk” modifications. The fractions from the ERLIC column were desalted prior to C-18-reversed phase (RP) LC-MS/MS analysis. Between 47-100% of the peptides with more than one phosphate or sialyl- residue or with a mannose-6 phosphate group were not retained by a C-18 cartridge but were retained by a cartridge of porous graphitic carbon. This finding implies that the abundance of such peptides may have been significantly underestimated in some past studies.
HostingRepository	PRIDE
AnnounceDate	2021-06-01
AnnouncementXML	Submission_2021-05-31_22:24:14.868.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Yusi Cui
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2020-12-07 23:34:02	ID requested
⏵ 1	2021-05-31 22:24:15	announced

Dataset with its publication pending

submitter keyword: ERLIC, PTM, Glyco, Phosphate, Peptide, Fraction

Lingjun Li
contact affiliation	1Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706 USA 2School of Pharmacy, University of Wisconsin-Madison, 777 Highland Ave., Madison, WI 53705 USA
contact email	lingjun.li@wisc.edu
lab head
Yusi Cui
contact affiliation	UW-Madison
contact email	cui42@wisc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022988
     1. Label: PRIDE project
     2. Name: Counterion Optimization Dramatically Improves Selectivity for Phosphopeptides and Glycopeptides in Electrostatic Repulsion-Hydrophilic Interaction Chromatography (ERLIC)