PhD Scientific Days 2024
Budapest, 9-10 July 2024
Poster Session R - Pharmaceutical Sciences and Health Technologies 2.
Identification of Glycosaminoglycan-Linker Glycopeptides in Proteoglycan Decorin Using HPLC-MS
Előadó neve
Ms. Dessidianti, Rachma
Neptun code
AO0ZTC
Előadó munkahelye
Semmelweis University Doctoral School, Budapest, Hungary
Előadó telefonszáma
+36205095164
Előadó e-mail címe
rachma.nur@phd.semmelweis.hu
Az előadás címe
Identification of Glycosaminoglycan-Linker Glycopeptides in Proteoglycan Decorin Using HPLC-MS
Szerző(k) neve és munkahelye
Rachma Dessidianti1, Mirjam Balbisi1, Lilla Turiák2
1: (1) MTA-TTK Lendület (Momentum) Glycan Biomarker Research Group, HUN-REN Research Centre for Natural Sciences, Budapest, Hungary; (2) Semmelweis University Doctoral School, Budapest, Hungary
2: MTA-TTK Lendület (Momentum) Glycan Biomarker Research Group, HUN-REN Research Centre for Natural Sciences, Budapest, Hungary
Bemutatás módja
Poszter
Szekció
Poster Session R - Pharmaceutical Sciences and Health Technologies 2.
Language of the presentation
English
Preferred session
Pharmaceutical Sciences and Health Technologies
Összefoglaló szövege
Introduction: Proteoglycans are generally composed of a core protein containing one or more covalently linked glycosaminoglycan (GAG) chains via a tetrasaccharide unit called the linker region. Proteoglycans play roles in normal and cancer cell physiology, with dysregulated expression contributing to cancer metastasis. The field of glycoproteomics focuses on the study of glycoproteins which also involves the identification and characterization of glycosaminoglycan-linker glycopeptides. In order to have a better understanding of the roles of glycoproteins in health and disease, it is essential to have a profound comprehension of the proteoglycan site-specific glycosylation alterations that occur throughout both normal and pathological processes.
Aims: Our aim is to develop a workflow to efficiently enrich and identify the glycosaminoglycan-linker glycopeptides that are present in decorin, a small leucine-rich proteoglycan, and to apply the workflow to more complex biological samples.
Methods: Decorin was sequentially digested using Lys-C/trypsin and chondroitinase ABC enzymes. Enrichment of glycosaminoglycan-linker glycopeptides was accomplished with the use of 10 kDa centrifugal filter membranes. Liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was utilized to identify the glycosaminoglycan-linker glycopeptides.
Results: Based on the extracted ion chromatograms, the presence of three diagnostic oxonium ions, [ΔHexAGalNAc]+ (m/z = 362.11), [GlcAGalNAc]+ (m/z = 380.12), and [GalNAc]+ (m/z = 204.09), could be identified indicating that the fraction contained chondroitin sulfate-linker glycopeptides. The exact structure of the glycopeptides could not be identified using Byonic software as tandem mass spectrometry conditions need further optimization.
Conclusions: The utilization of Lys-C/trypsin and chondroitinase ABC enzymes in conjunction with a 10 kDa centrifugal filter membrane show promising results for the enrichment of chondroitin sulfate-linker glycopeptides.
Further Studies: A comparative analysis will be conducted using different enrichment methods for glycosaminoglycan-linker glycopeptides identification using decorin standard.
Acknowledgments: Support of the Lendület (Momentum) Program of the Hungarian Academy of Sciences is acknowledged. The Authors are grateful to Markus Ralser for donating the nanoAcquity HPLC to the group.
University
Semmelweis University
Supervisor
Lilla Turiák
Publication of my abstract
I give consent to the publication of my abstract on the website of the congress.
Kind
Szabad
Status
elfogadva
Accepted presentation method
poszter
Előadás fájl jóváhagyás
nem rendelkezett róla
Előadó
8048
Start
16:15
End
16:18