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An enzyme-based screening system for the rapid assessment of protein N-glycosylation efficiency in yeast

Frey, Alexander D. ; Aebi, Markus

In: Glycobiology, 2015, vol. 25, no. 3, p. 252-257

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    Titel
    • An enzyme-based screening system for the rapid assessment of protein N-glycosylation efficiency in yeast
    Autor
    • Frey, Alexander D.. Department of Biotechnology and Chemical Technology, Aalto University, Aalto FI-00076, Finland
    • Aebi, Markus. Institute of Microbiology, ETH Zürich, Zürich CH-8093, Switzerland
    Dokumententyp
    • Postprint
    Sprache
    • Englisch
    Veröffentlicht in
    • Glycobiology, 2015, vol. 25, no. 3, p. 252-257. Oxford University Press
    Andere elektronische Ausgabe
    OAI-PMH-ID
    • oai:doc.rero.ch:302184
    Summary
    N-Glycosylation efficiency is a key parameter when studying components of the protein N-glycosylation pathway, but was recently also recognized as an important factor in the production of glycosylated proteins. We have developed a novel assay to quantify N-glycosylation efficiency of proteins. This assay is based on the secreted activity of yeast acid phosphatase, the proper folding and hence secretion of which is strongly dependent on its N-glycosylation status. The results show that the reporter yields a quantitative measure for protein N-glycosylation in yeast, which is in good agreement with classically used assay based on protein migration patterns on SDS-PAGE. However, the assay is less laborious and is adaptable to high-throughput screening approaches as exemplified