Metabolic engineering of glycoproteins (B05)

• Subject Area: Biological and Biomimetic Chemistry
• Term: from 2012 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 189682160

Glycosylation is a complex form of posttranslational modification that is involved in the regulation of many aspects of protein function. In this project, metabolic glycoengineering is used to incorporate chemical reporter groups into carbohydrate structures of glycoproteins. Subsequently, the glycoproteins can be labeled with a probe (fluorescent dye, biotin) through a bioorthogonal ligation reaction enabling their visualization or isolation. In the past, we established the inverse-electron-demand Diels-Alder reaction as a suitable bioorthogonal ligation reaction for this purpose employing terminal alkenes, cyclopropenes, and norbornenes as chemical reporter groups. We will now develop new carbohydrate probes that are incorporated with higher efficiency. With the new probes, we will study protein glycosylation in living cells. Specifically, we will investigate dynamic O-GlcNAcylation of the kinesin Kif18A. Another goal is the development of a chemical method to detect and distinguish protein O-GlcNAcylation and phosphorylation through selective elimination of GlcNAc and phosphate, respectively, followed by Michael addition of a label and proteomic analysis.

• DFG Programme: Collaborative Research Centres
• Subproject of: SFB 969: Chemical and Biological Principles of Cellular Proteostasis
• Applicant Institution: Universität Konstanz
• Project Head: Professor Dr. Valentin Wittmann