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Semisynthesis of Functional Glycosylphosphatidylinositol-Anchored Proteins |
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| Authors: | Dr Renée F Roller Dr Ankita Malik Dr Maria A Carillo Dr Monika Garg Antonella Rella Marie-Kristin Raulf Prof Dr Bernd Lepenies Prof Dr Peter H Seeberger Dr Daniel Varón Silva |
| Institution: | 1. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany;2. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany
Institute of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany These authors contributed equally to this work.;3. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany;4. Immunology Unit and Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany Institute for Parasitology, Center for infection Medicine, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany;5. Immunology Unit and Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany |
| Abstract: | Glypiation is a common posttranslational modification of eukaryotic proteins involving the attachment of a glycosylphosphatidylinositol (GPI) glycolipid. GPIs contain a conserved phosphoglycan that is modified in a cell- and tissue-specific manner. GPI complexity suggests roles in biological processes and effects on the attached protein, but the difficulties to get homogeneous material have hindered studies. We disclose a one-pot intein-mediated ligation (OPL) to obtain GPI-anchored proteins. The strategy enables the glypiation of folded and denatured proteins with a natural linkage to the glycolipid. Using the strategy, glypiated eGFP, Thy1, and the Plasmodium berghei protein MSP119 were prepared. Glypiation did not alter the structure of eGFP and MSP119 proteins in solution, but it induced a strong pro-inflammatory response in vitro. The strategy provides access to glypiated proteins to elucidate the activity of this modification and for use as vaccine candidates against parasitic infections. |
| Keywords: | GPI anchor glycoproteins glypiation protein modifications protein semisynthesis |