Active RNase glycoprotein from three pieces: The glycoprotein enzyme ribonuclease C, which contains a complex saccharide N-glycan, was synthesized by sequential native chemical ligation. An optimized ligation and isolation protocol allowed the efficient assembly and refolding of the 124 amino acid enzyme.
Seven in one blow: The efficient formation of mixed disulfides on the thiol-rich fusion protein A followed by subsequent intein cleavage gave the fragment B with all seven cysteines protected against oxidation. The native chemical ligation of B with synthetic glycopeptide thioesters provides glycoproteins.
Sieben auf einen Streich: Durch die effiziente Bildung von gemischten Disulfiden an dem thiolreichen Fusionsprotein A konnte nach Inteinabspaltung das Cys‐Fragment B mit sieben oxidationsgeschützten Cysteinen erhalten werden. Die native chemische Ligation von B mit synthetischen Glycopeptidthioestern ergibt Glycoproteine.
Aktives RNase‐Glycoprotein aus drei Segmenten: Das Glycoprotein‐Enzym Ribonuclease C mit einem komplexen nonasaccharidischen N‐Glycan wurde durch sequenzielle native chemische Ligation synthetisiert. Durch optimierte Ligations‐ und Isolierungsbedingungen konnte das aus 124 Aminosäuren bestehende Enzym effizient aufgebaut und zurückgefaltet werden.
Enzymes U 1100 Semisynthesis of a Homogeneous Glycoprotein Enzyme: Ribonuclease C. Part 2. -(PIONTEK, C.; SILVA, D. V.; HEINLEIN, C.; POEHNER, C.; MEZZATO, S.; RING, P.; MARTIN, A.; SCHMID, F. X.; UNVERZAGT*, C.; Angew.
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