Exploiting Diels-Alder Reactions with Inverse Electron Demand for Bioorthogonal Labeling of Alkene-Substituted Fucosylated Glycoproteins in Cells

Glycosylation is an ubiquitous form of posttranslational modification that has been estimated to occur on more than 50% of the proteins from eukaryotic cells. Among the nine monosaccharides found in the glycoproteins of vertebrates, fucose is distinguished by its occurrence in numerous epitopes associated with cell-cell interactions and regulation of protein function. Fucose is also contained in a number of tumor-associated antigens that are upregulated in cancerous tissues and, therefore, have been suggested to act as cancer biomarkers. Profiling fucosylated glycoproteins is a possible strategy for the detection of these biomarkers.

The aim of this project is the application of the Diels-Alder reaction with inverse electron demand (DARinv) of terminal alkenes as a new bioorthogonal ligation technique for the metabolic labeling of fucosylated glycoproteins in cells. Cells will be cultivated in the presence of synthetic fucose derivatives containing terminal alkenes that function as dienophiles. After metabolic incorporation into cellular glycoproteins, the dienophiles can be chemoselectively addressed by a DARinv with tetrazine derivatives carrying a detectable probe (fluorescence dye, biotin). A major advantage of the DARinv is that it is orthogonal to click chemistry. This will allow for selective staining of two sugars (one by DARinv, the other by click chemistry) independently and in presence of the other. The methods that will be developed in this project enable functional analyses of protein fucosylation in whole cells.

Prof. Dr. Valentin Wittmann
Universität Konstanz

Tel.: +49 7531 88 - 4572
Fax: +49 7531 88 - 4573

Email Prof. Wittmann

Publications within the SPP 1623 project

J.E.G.A. Dold, V. Wittmann
ChemBioChem2021, 22, 1243-1251
Metabolic Glycoengineering with Azide- and Alkene-Modified Hexosamines: Quantification of Sialic Acid Levels
Link to the article

L.M. Haiber, M. Kufleitner, V. Wittmann
Front. Chem. 2021, 9, 654932
Application of the Inverse-Electron-Demand Diels-Alder Reaction for Metabolic Glycoengineering
Link to the article

M.J. Schöwe, O. Keiper, C. Unverzagt, V. Wittmann
Chem. Eur. J. 2019, 25, 15759-15764
A Tripeptide Approach to the Solid-Phase Synthesis of Peptide Thioacids and N-Glycopeptides
Link to the article

V. F. Schart, J. Hassenrück, A.-K. Späte, J. E. G. A. Dold, R. Fahrner, V. Wittmann
ChemBioChem2019, 20, 166-171
Triple Orthogonal Labeling of Glycans Applying Photoclick Chemistry
Link to the article

J.E.G.A. Dold, J. Pfotzer, A.-K. Späte, V. Wittmann
ChemBioChem 2017, 18(13), 1242-1250
Dienophile-Modified Mannosamine Derivatives for Metabolic Labeling of Sialic Acids: A Comparative Study
Link to the article

A.-K. Späte, J.E.G.A. Dold, E. Batroff, V.F. Schart, D.E. Wieland, O.R. Baudendistel, V. Wittmann
ChemBioChem 2016, 17, 1274-1383
Exploring the Potential of Norbornene-Modified Mannosamine Derivatives for Metabolic Glycoengineering.
Link to the article

F. Doll, A. Buntz, A.-K. Späte, V.F. Schart, A. Timper, W. Schrimpf, C.R. Hauck, A. Zumbusch, V. Wittmann
Angew. Chem. Int. Ed. 2016, 55, 2262-2266
Visualization of Protein-Specific Glycosylation inside Living Cells.
Link to the article

K. Rohmer, J. Mannuthodikayil, V. Wittmann
Isr. J. Chem. 2015, 55( 3-4), 437-446
Application of the Thioacid-Azide Ligation (TAL) for the Preparation of Glycosylated and Fluorescently Labeled Amino Acids
Link to the article

A.-K. Späte, V.F. Schart, S. Schöllkopf, A. Niederwieser, V. Wittmann
Chem. Eur. J. 2014, 20 (50), 16502-16508
Terminal Alkenes as Versatile Chemical Reporter Groups for Metabolic Oligosaccharide Engineering
Link to the article

A.-K. Späte, H. Bußkamp, A. Niederwieser, V. F. Schart, A. Marx, V. Wittmann
Bioconjug Chem.2014, 25, 147-154
Rapid Labeling of Metabolically Engineered Cell-Surface Glycoconjugates with a Carbamate-Linked Cyclopropene Reporter
Link to the article