Semisynthesis of Functional Glycosylphosphatidylinositol‐Anchored Proteins |
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Authors: | Rene F Roller Ankita Malik Maria A Carillo Monika Garg Antonella Rella Marie‐Kristin Raulf Bernd Lepenies Peter H Seeberger Daniel Varn Silva |
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Institution: | Renée F. Roller,Ankita Malik,Maria A. Carillo,Monika Garg,Antonella Rella,Marie‐Kristin Raulf,Bernd Lepenies,Peter H. Seeberger,Daniel Varón Silva |
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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. |
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Keywords: | GPI anchor glycoproteins glypiation protein modifications protein semisynthesis |
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