EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins

Erik J. Liu, Andrew Sinclair, Andrew J. Keefe, Brent Nannenga, Brandon L. Coyle, François Baneyx, Shaoyi Jiang

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3357-3361
Number of pages5
JournalBiomacromolecules
Volume16
Issue number10
DOIs
StatePublished - Oct 12 2015
Externally publishedYes

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Peptides
Proteins
Biosynthesis
Bioactivity
Polyethylene glycols
Lysine
Glutamic Acid
Polymers
Fusion reactions
Enzymes
Stabilization
Salts
Transmission electron microscopy
Acids
Temperature

ASJC Scopus subject areas

  • Bioengineering
  • Materials Chemistry
  • Polymers and Plastics
  • Biomaterials

Cite this

Liu, E. J., Sinclair, A., Keefe, A. J., Nannenga, B., Coyle, B. L., Baneyx, F., & Jiang, S. (2015). EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins. Biomacromolecules, 16(10), 3357-3361. https://doi.org/10.1021/acs.biomac.5b01031

EKylation : Addition of an Alternating-Charge Peptide Stabilizes Proteins. / Liu, Erik J.; Sinclair, Andrew; Keefe, Andrew J.; Nannenga, Brent; Coyle, Brandon L.; Baneyx, François; Jiang, Shaoyi.

In: Biomacromolecules, Vol. 16, No. 10, 12.10.2015, p. 3357-3361.

Research output: Contribution to journalArticle

Liu, EJ, Sinclair, A, Keefe, AJ, Nannenga, B, Coyle, BL, Baneyx, F & Jiang, S 2015, 'EKylation: Addition of an Alternating-Charge Peptide Stabilizes Proteins', Biomacromolecules, vol. 16, no. 10, pp. 3357-3361. https://doi.org/10.1021/acs.biomac.5b01031
Liu, Erik J. ; Sinclair, Andrew ; Keefe, Andrew J. ; Nannenga, Brent ; Coyle, Brandon L. ; Baneyx, François ; Jiang, Shaoyi. / EKylation : Addition of an Alternating-Charge Peptide Stabilizes Proteins. In: Biomacromolecules. 2015 ; Vol. 16, No. 10. pp. 3357-3361.
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