Thesis supervisor: Csaba Matta
Location of studies (in Hungarian): University of Debrecen, Faculty of Medicine, Department of Anatomy, Histology & Embryology Abbreviation of location of studies: DEASZ
Description of the research topic:
Proteins that are associated with the plasma membrane constitute the surfaceome. The aim of the current PhD project is to test a refined method for more selective surface protein labelling and enrichment, which will hopefully minimise the false positive protein IDs in samples enriched for surface proteins. We are going to use this method on cartilage progenitor cells (CPCs) and chondrocytes isolated from human articular cartilage (both healthy and osteoarthritic). The methodology that the PhD candidate is expected to implement, fine-tune and apply relies on cell lysates. Following reduction and alkylation, proteins will be digested using Trypsin/Lys-C protease mix. Following digestion, the glycan chains of glycopeptides will be oxidized using sodium-metaperiodate, and the resulting oxidized glycopeptides will be captured on hydrazide beads. After thorough washes, N-glycopeptides will be enzymatically deglycosylated and eluted using PNGase F. Since the PNGase F-cleavage of glycan chains bound to asparagine residues results in the conversion of asparagine to aspartic acid, downstream bioinformatic analysis will be performed by searching for these deamidation events. This way, we are expecting a much higher proportion of the identified proteins to be localized to the plasma membrane. The proposed research will have significant impact across a number of different fields. Identifying novel biomarkers during articular cartilage formation can lead to the development of drugs, or the identification of external cues that reactivate the embryonic developmental mechanisms. This could lead to novel repair strategies to fully restore joint function; by systemically targeting specific biomarkers we may be able to slow down disease progression and/or enhance intrinsic cartilage repair.
Required language skills: English intermediate level Number of students who can be accepted: 1
Deadline for application: 2024-05-15
2024. IV. 17. ODT ülés Az ODT következő ülésére 2024. június 14-én, pénteken 10.00 órakor kerül sor a Semmelweis Egyetem Szenátusi termében (Bp. Üllői út 26. I. emelet).
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