EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins

For nearly 40 years, therapeutic proteins have been stabilized by chemical conjugation of polyethylene glycol (PEG), but recently zwitterionic materials have proved to be a more effective substitute. In this work, we demonstrate that genetic fusion of alternating-charge extensions consisting of anio...

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Bibliographic Details
Published in:Biomacromolecules 2015-10, Vol.16 (10), p.3357-3361
Main Authors: Liu, Erik J, Sinclair, Andrew, Keefe, Andrew J, Nannenga, Brent L, Coyle, Brandon L, Baneyx, François, Jiang, Shaoyi
Format: Article
Language:English
Subjects:
Anions
Cations
Glutamic Acid - chemistry
Kinetics
Lysine - chemistry
Peptides - chemistry
Proteins - chemistry
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Summary:For nearly 40 years, therapeutic proteins have been stabilized by chemical conjugation of polyethylene glycol (PEG), but recently zwitterionic materials have proved to be a more effective substitute. In this work, we demonstrate that genetic fusion of alternating-charge extensions consisting of anionic glutamic acid (E) and cationic lysine (K) is an effective strategy for protein stabilization. This bioinspired “EKylation” method not only confers the stabilizing benefits of poly­(zwitterions) but also allows for rapid biosynthesis of target constructs. Poly­(EK) peptides of different predetermined lengths were appended to the C-terminus of a native β-lactamase and its destabilized TEM-19 mutant. The EK-modified enzymes retained biological activity and exhibited increased stability to environmental stressors such as high temperature and high-salt solutions. This one-step strategy provides a broadly applicable alternative to synthetic polymer conjugation that is biocompatible and degradable.
ISSN:1525-7797
1526-4602
DOI:10.1021/acs.biomac.5b01031