PXD047065

PXD047065 is an original dataset announced via ProteomeXchange.

Title	TIHAMAC: Streamlined Tandem IMAC-HILIC Workflow for Simultaneous and High-Throughput Plant Phosphoproteomics and N-glycoproteomics Analysis
Description	Protein post-translational modifications (PTMs) are pivotal molecular events in plant cells, encompassing processes such as protein folding and enzyme activation. Among these PTMs, N-glycosylation and phosphorylation hold particular importance, as they play essential roles in regulating plant growth and stress responses. The intricate PTM landscape is exemplified by the identification of multiple plant membrane-localized proteins subject to both phosphorylation and N-glycosylation, hinting at potential interplay influencing protein functionality. To investigate the crosstalk between phosphorylation and glycosylation in plant signaling, immobilized metal affinity chromatography (IMAC) and hydrophilic interaction chromatography (HILIC) have been primarily utilized for large-scale phosphopeptide and glycopeptide enrichment, respectively. In this work, we introduce a novel streamlined tandem S-Trap-IMAC-HILIC strategy, termed TIMAHAC, to enable serial enrichment of phosphopeptides and glycopeptides. This approach successfully integrates the two methods through tandem tip enrichment, eliminating the need for buffer exchange and sample transfer. As a result, it significantly minimizes sample loss during serial enrichment steps, and streamlines experimental procedures, ultimately saving time. We demonstrated the efficiency of the TIHAMAC strategy in the glycoproteomics and phosphoproteomics analyses using 200 μg of Arabidopsis digests. Up to 1,954 N-glycopeptides and 11,255 phosphopeptides were identified by timsTOF HT. Remarkably, 69% of N-glycopeptides were consistently quantified with a coefficient of variation less than 20%. Furthermore, more than one-third identified glycoproteins exhibited phosphorylation modifications. These results suggest that the applicability of the TIMAHAC method for studying the potential crosstalk of N-glycoproteome and phosphoproteome within complex plant signaling networks.
HostingRepository	PRIDE
AnnounceDate	2024-04-03
AnnouncementXML	Submission_2024-04-03_09:38:51.148.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Chuan-Chih Hsu
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	phosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	autoflex

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-19 05:10:26	ID requested
⏵ 1	2024-04-03 09:38:51	announced

Dataset with its publication pending

submitter keyword: N-glycoproteomics, HILIC, Arabidopsis thaliana,IMAC, ABA signaling, phosphoproteomics

Chuan-Chih Hsu
contact affiliation	Institute of Plant and Microbial Biology, Academia Sinica
contact email	cchsu@gate.sinica.edu.tw
lab head
Chuan-Chih Hsu
contact affiliation	Academia Sinica
contact email	without1102@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD047065
     1. Label: PRIDE project
     2. Name: TIHAMAC: Streamlined Tandem IMAC-HILIC Workflow for Simultaneous and High-Throughput Plant Phosphoproteomics and N-glycoproteomics Analysis