PXD024196

PXD024196 is an original dataset announced via ProteomeXchange.

Title	Complementary Proteome and Glycoproteome Access Revealed Through Comparative Analysis of Reversed Phase and Porous Graphitic Carbon Chromatography
Description	Aided by improvements in instrumental and analytical techniques, rigorous connections have been drawn between glycoprotein expression, modification heterogeneity, and a variety of human illnesses. These illnesses, such as various forms of cancer, are often associated with poor prognoses, prompting the need for more comprehensive characterization of the glycoproteome. Instrumental advancements, optimal and nascent dissociation techniques, and computational strategies present the largest area of glycoprotein profiling innovation, often neglecting potential benefits provided by alternative chromatography techniques. Porous graphitic carbon (PGC) represents a separation regime that has gained significant interest in glycomics research due to its mobile phase flexibility, increased retention of polar analytes and improved structural elucidation at higher temperatures. PGC has yet to be systematically compared against or in tandem with standard reversed phase liquid chromatography (RPLC) in high-throughput bottom-up glycoproteomics experiments, leaving the potential benefits unexplored. Performing comparative analysis of single and biphasic separation regimes at a range of column temperatures illustrates the advantages for each method. PGC separation is shown to selectively retain shorter, more hydrophilic glycopeptide species, imparting higher average charge, and exhibiting greater microheterogeneity coverage for identified glycosites. Additionally, we demonstrate that liquid-phase separation of glycopeptide isomers may be achieved when in both single and biphasic PGC separation, providing a means towards facile, multiplex glycopeptide characterization. Beyond this, we demonstrate how utilization of multiple separation regimes and column temperatures can aid in profiling the glycoproteome in tumorigenic and aggressive prostate cancer cells.
HostingRepository	PRIDE
AnnounceDate	2022-02-14
AnnouncementXML	Submission_2022-02-14_01:30:23.738.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD024196
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Daniel Delafield
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-02-16 00:23:47	ID requested
⏵ 1	2022-02-14 01:30:24	announced

Delafield DG, Miles HN, Liu Y, Ricke WA, Li L, Complementary proteome and glycoproteome access revealed through comparative analysis of reversed phase and porous graphitic carbon chromatography. Anal Bioanal Chem, 414(18):5461-5472(2022) [pubmed]

submitter keyword: Glycoproteomics, glycopeptides, proteomics, mass spectrometry

Lingjun Li
contact affiliation	University of Wisconsin-Madison
contact email	lingjun.li@wisc.edu
lab head
Daniel Delafield
contact affiliation	University of Wisconsin - Madison
contact email	delafield@wisc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD024196
     1. Label: PRIDE project
     2. Name: Complementary Proteome and Glycoproteome Access Revealed Through Comparative Analysis of Reversed Phase and Porous Graphitic Carbon Chromatography