Project Details
Projekt
Unravelling the correlation between different ER-based types of glycosylation via glyco/proteo-analytics
Applicant
Dr. Erdmann Rapp
Subject Area
Cell Biology
Biochemistry
Biochemistry
Term
from 2017 to 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 289991887
Protein glycosylation, i.e. the covalent attachment of single monosaccharides or complex oligosaccharides to specific amino acids within a peptide sequence, is a co- and post-translational modification conserved across all domains of life (bacteria, archaea, eukaryota). Located on the majority of secreted or membrane-bound proteins, glycans are involved in virtually every cellular event, and essential for vertebrate development and physiology. N-glycosylation, the most common form of glycosylation, is initiated at the endoplasmic reticulum translocon (ER) and requires nucleotide sugars and dolichol-linked precursors (e.g. guanosine diphosphate mannose (GDP-Man) and dolichol monophosphate mannose (Dol-P-Man)) for the early stages of synthesis. Two other ER-resident forms of glycosylation, C-mannosylation and O-mannosylation, also depend on Dol-P-Man for their initial synthesis. Dependencies, compensatory effects, and the competition for the common mannose substrates and acceptor proteins of these three dolichol-based glycosylation pathways are largely unknown, but of high relevance to understand glycosylation related diseases and disorders, such as infectious and autoimmune diseases, cancer, and congenital disorders of glycosylation (CDG). To unravel their interplay, we intend to study N-glycosylation, C-mannosylation, O-mannosylation in different model systems (cell lines, patient cells, and fish lines), deficient for either of the three pathways, as well as model proteins produced in these systems, e.g. E-cadherin derived from POMT2-/-/POMGnT1-/- HEK293 cells. To obtain a holistic view on resulting glycosylation alterations, these model systems will be analysed with respect to changes in glycan macro- and microheterogeneity using our comprehensive and state-of-the art glyco/proteo-analytical toolbox.
DFG Programme
Research Units
