EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins

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• 作者：Erik J. Liu ; Andrew Sinclair ; Andrew J. Keefe ; Brent L. Nannenga ; Brandon L. Coyle ; Fran莽ois Baneyx ; Shaoyi Jiang
• 刊名：Biomacromolecules
• 出版年：2015
• 出版时间：October 12, 2015
• 年：2015
• 卷：16
• 期：10
• 页码：3357-3361
• 全文大小：339K
• ISSN：1526-4602

文摘

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 鈥淓Kylation鈥?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 尾-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.