Nonenzymatic synthesis of anomerically pure, mannosyl-based molecular probes for scramblase identification studies

Giovanni Picca; Markus Probst; Simon M Langenegger; Oleg Khorev; Peter Bütikofer; Anant K Menon; Robert Häner

doi:10.3762/bjoc.16.145

Nonenzymatic synthesis of anomerically pure, mannosyl-based molecular probes for scramblase identification studies

Beilstein J Org Chem. 2020 Jul 20:16:1732-1739. doi: 10.3762/bjoc.16.145. eCollection 2020.

Authors

Giovanni Picca¹, Markus Probst¹, Simon M Langenegger¹, Oleg Khorev¹, Peter Bütikofer², Anant K Menon³, Robert Häner¹

Affiliations

¹ Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
² Institute of Biochemistry and Molecular Medicine, University of Bern, Bühlstrasse 28, 3012 Bern, Switzerland.
³ Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, 10065 New York, United States of America.

Abstract

The chemical synthesis of molecular probes to identify and study membrane proteins involved in the biological pathway of protein glycosylation is described. Two short-chain glycolipid analogs that mimic the naturally occurring substrate mannosyl phosphoryl dolichol exhibit either photoreactive and clickable properties or allow the use of a fluorescence readout. Both probes consist of a hydrophilic mannose headgroup that is linked to a citronellol derivative via a phosphodiester bridge. Moreover, a novel phosphoramidite chemistry-based method offers a straightforward approach for the non-enzymatic incorporation of the saccharide moiety in an anomerically pure form.

Keywords: carbohydrates; citronellol; phosphoramidite; photoclickable glycolipid analogs; scramblase.

Grants and funding

Financial support by the Swiss National Science Foundation (SNSF) is gratefully acknowledged: This research was funded by the Sinergia Project: Molecular identification of lipid transporters for protein glycosylation, Grant CRSII5_170923.