Enzymatically controlled release of proteins and peptides: A promising, alternative secretion approach
Daniel Ivanusic, Gregor Zipf, Josef Maier, Hubert Bernauer, Norbert Bannert
TL;DR
This paper introduces ENCOREP, a new method to release glycosylated proteins from cells, enabling the production of hard-to-obtain glycopeptides for research.
Contribution
ENCOREP offers a novel, enzyme-driven approach to release glycosylated proteins from the cell surface, bypassing traditional secretion methods.
Findings
ENCOREP successfully produced glycosylated peptides from the CD63 protein.
The method uses an engineered protease for continuous release of extracellular loops.
Released proteins are suitable for functional and biochemical assays.
Abstract
The biological activity of many proteins is influenced by glycosylation, underscoring the essential role of the glycosylation process. However, no established recombinant technique currently enables the controlled shedding of glycosylated extracellular loops from transmembrane proteins. Here, we describe enzymatically controlled release of proteins and peptides (ENCOREP), a strategy that enables in situ expression at the cell surface and protease-mediated release, followed by collection from the cell culture medium. Using ENCOREP, we achieved the production of glycosylated peptides. These glycopeptides, derived from the large extracellular loop of the highly glycosylated CD63, illustrate the type of targets that are otherwise inaccessible with current methods but can be readily obtained using ENCOREP. Overall, ENCOREP provides a rapid and reliable approach to obtain glycosylated…
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Taxonomy
TopicsAntimicrobial Peptides and Activities · Biochemical and Structural Characterization · RNA Interference and Gene Delivery