Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Development and validation of a method for analyzing the sialylated glycopeptides of recombinant erythropoietin in urine using LC-HRMS

ABSTRACT: 1. Profiling of sialylated glycopeptides from rEPO was performed via LC–HRMS 2. LC–HRMS methods for analyzing sialylated glycopeptide in urine samples were developed 3. The method was validated and applied to detection of rEPO biosimilars in urine

INSTRUMENT(S): Orbitrap Exploris 240

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Urine

SUBMITTER: YOONDAM SEO

LAB HEAD: Yoondam, Seo

PROVIDER: PXD038932 | Pride | 2023-05-10

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	210806_BRP_1mg_Glu_C_Trypsin_TiO2_bulk_fullms_ddms.raw	Raw
	210808_BRP_1mg_Glu_C_Trypsin_TiO2_fullms_ddms_final.xlsx	Xlsx

Items per page:

1 - 2 of 2

Publications

Development and validation of a method for analyzing the sialylated glycopeptides of recombinant erythropoietin in urine using LC-HRMS.

Seo Yoondam Y Park Jisoo J Lee Hyeon-Jeong HJ Kim Minyoung M Kang Inseon I Son Junghyun J Oh Min-Kyu MK Min Hophil H

Scientific reports 20230308 1

Erythropoietin (EPO) is a glycoprotein hormone that stimulates red blood cell production. It is produced naturally in the body and is used to treat patients with anemia. Recombinant EPO (rEPO) is used illicitly in sports to improve performance by increasing the blood's capacity to carry oxygen. The World Anti-Doping Agency has therefore prohibited the use of rEPO. In this study, we developed a bottom-up mass spectrometric method for profiling the site-specific N-glycosylation of rEPO. We reveale ...[more]

PMID: 36890204

Similar Datasets

Site-specific glycosylation of donkey milk lactoferrin investigated by High Resolution Mass Spectrometry

Project description:A comprehensive glycosylation profile of donkey lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. The results obtained allowed the identification of 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. Altogether, the N-glycan structures determined revealed that in donkey milk lactoferrin most of the N-glycans identified are neutral complex/hybrid. Actually, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. In addition, 2 high mannose N-glycans, 4 sialylated fucosylated complex/hybrid N-glycans and 6 sialylated non-fucosylatedN-glycans, one of which containing N-glycolylneuramin acid (Neu5Gc), were found. A comparison of the glycosylation profile of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported.

2016-08-25 | PXD004289 | Pride

Characterization of signaling pathways associated with pancreatic beta-cell adaptive flexibility in compensation of obesity-linked diabetes in db/db miceCharacterization of signaling pathways associated with pancreatic β-cell adaptive flexibility in compensation of obesity-linked diabetes in db/db mice

Project description:To characterize the signaling mechanisms underlying beta-cell dysfunction in db/db mice and their return to a more “normal” beta-cell phenotype, we applied a mass spectrometry-based quantitative phosphophoproteomics and sialiomics(sialylated N-linked glycopeptides) approach to normal and db/db beta-cells from hyperglycemic and euglycemic environments.

2020-05-19 | PXD017469 | Pride

Cellular signaling underlying GSIS in pancreatic β-cells

Project description:In the present study, we investigated signaling pathways associated with GSIS in freshly isolated rat Islets of Langerhans. Specifically, our aim was to reveal which phosphosites and N-linked sialylated glycosylation sites that were reversible regulated upon brief glucose stimulation and link these to cellular signaling pathways using bioinformatics tools.

2017-11-14 | PXD007689 | Pride

Synaptic transmission induces site specific changes in sialylation on N-linked glycoproteins in nerve terminals

Project description:Synaptic transmission is an ultra-fast process that aims at transducing a signal from one neuron to another neuron or cell by the release of neurotransmitters. This process requires rapid response which can be transmitted through fast molecular switches such as post-translational modifications of proteins. We aimed at investigating the potential importance in dynamic modulation of sialic acids on N-linked glycoproteins that are essential for the synaptic transmission process. For this, we characterize site-specific alteration in sialylated N-linked glycosylation in the active zone of rat nerve terminals (synaptosomes) after depolarization by using quantitative sialiomics.

2020-06-12 | PXD016230 | Pride

Solid-phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for comprehensive characterization of glycoproteins

Project description:A novel chemoenzymatic method termed solid phase extraction of N-linked Glycans And Glycosite-containing peptides (NGAG) for the simultaneous analysis of N-glycans, glycosite-containing peptides, and intact N-glycopeptides with site-specific glycosylation information.

2015-11-23 | PXD001571 | Pride

Characterization of sialylated N- and O-glycopeptides from complex proteomes on an Orbitrap Fusion Tribrid mass spectrometer by Isotope Targeted Glycoproteomics (IsoTaG). Application to azido and alkynyl sugars

Project description:Protein glycosylation is a post-translational modification (PTM) responsible for many aspects of proteomic diversity and biological regulation. Correlation of the intact glycoform to the protein attachment site is a critical step to assign functional roles to specific glycoproteins. Isotope targeted glycoproteomics (IsoTaG) is a mass-independent mass spectrometry method to characterize intact, metabolically labeled glycopeptides from complex proteomes. IsoTaG was applied to conditioned media from PC-3 cells labeled with alkynyl or azido sugars to reveal the sialylated glycoproteome. Analysis on an Orbitrap Fusion Tribrid mass spectrometer resulted in characterization of 699 intact glycopeptides from 192 glycoproteins. These intact glycopeptides represent a total of eight sialylated glycoforms across 126 Nand 576 O-glycopeptides. IsoTaG is an effective platform for identification of intact glycopeptides labeled by alkynyl or azido-glycans and will facilitate further studies on the role of the glycoproteome in biology.

2017-02-22 | PXD004302 | Pride

Comprehensive analysis of the proteome, phosphoproteome and N-linked sialylated glycoproteome of C2C12 myoblasts

Project description:C2C12 myoblast is a model that has been used extensively to study the process of skeletal muscle differentiation. Proteomics has advanced our understanding of skeletal muscle biology and the process of myogenesis. However, there is still no deep coverage of C2C12 myoblast proteome, which is important for the understanding of key drivers for the differentiation of skeletal muscle cells. Here, we conducted a multi-dimensional proteome profiling with TiSH strategy to get a comprehensive analysis of proteome, phosphoproteome and N-linked sialylated glycoproteome of C2C12 myoblasts. A total of 8313 protein groups were identified in C2C12 myoblasts, including 7827 protein groups from non-modified peptides, 3803 phosphoproteins and 977 formerly N-linked sialylated glycoproteins.

2020-11-27 | PXD011334 | Pride

Chronic low-dose ionising radiation rates affect the hippocampal phospho-proteome of synaptogenesis in an ApoE-/--Alzheimer mouse model over 300 days of exposure

Project description:Cognitive impairments are an important side effect after radiation exposure. Yet, it comes more and more into scientific focus that ionising radiation can resemble pathologies of neurodegenerative diseases such as Alzheimer´s. Here, we used a multidisciplinary approach to elucidate the effect of chronic low-dose irradiation exposure (1 mGy/day and 20 mGy/day) over 300 days (cumulative dose of 0.3 Gy and 6.0 Gy) on the murine ApoE-/- hippocampus as an Alzheimer´s model. By using mass spectrometry to quantify global expression levels of unmodified proteins, phospho-proteins and N-linked sialylated glycoproteins, we found nearly exclusive changes on the phospho-proteome at both doses. These proteins regulate signalling of synaptic plasticity and calcium-dependent signalling. We used synaptic plasticity-targeted transcriptomics, immunoblotting and ELISA to validate our findings and could identify that memory-related CREB signalling is reduced at both doses whereas Rac1-Cofilin signalling is only altered at 1 mGy/day. Interestingly, we noted a reduction in the number of activated microglia in the molecular layer of the hippocampus only at 1 mGy/day paralleled with reduced levels of TNFα mRNA and lipid peroxidation at this dose. Adult neurogenesis (Ki67, GFAP and NeuN as markers) and cell death (activated caspase-3) were not changed / initiated at both doses. Our analysis showed that several molecular targets induced by chronic low-dose radiation overlap with molecular targets of Alzheimer´s pathology. We suggest that chronic low-dose ionising radiation can be a potential confounder in Alzheimer´s disease.

2017-07-25 | PXD003969 | Pride

Optimization of O-GIG for O-glycopeptide Characterization with Sialic Acid Linkage Determination-Fig6

Project description:O-glycoprotein analysis has been historically challenging due, in part, to a dearth of available enzymes active in release of O-glycans. Moreover, chemical releasing methods, such as β-elimination/Michael addition are not specific to O-glycan release and can also eliminate phosphoryl substitutions. Both of these events leave behind deaminated serine and threonine and thus can lead to ambiguous structural conclusions. Recently, the O-protease OpeRATOR, derived from intestinal bacteria and expressed in E. coli, has become commercially available. Digestion of O-glycoprotein yields O-glycopeptides cleaved at the N-terminal end of serine/threonine with O-glycan remaining intact. The enzyme has broad substrate specificity and includes mammalian Cores 1-8. However, OpeRATOR is not fully active toward sialylated glycoproteins and therefore it is suggested that this acidic residue be removed prior to digestion, thus, sacrificing structural information. In this study, we investigated the performance of OpeRATOR under a range of conditions, including buffer selection, pH, sialic acid modification, and digestion temperature in order to optimize enzymatic activity with special emphasis on sialylated glycosites. Conditions derived in this work facilitate OpeRATOR digestion of fully sialylated O-glycopeptides mass tagged to identify the sialyl linkage, thus, facilitating analysis of these charged O-glycopeptides, which are often important in biological processes.

2020-07-28 | PXD020034 | Pride

Optimization of O-GIG for O-glycopeptide Characterization with Sialic Acid Linkage Determination-Fig3

2020-07-28 | PXD020048 | Pride