Per-glycosylation of the Surface-Accessible Lysines: One-Pot Aqueous Route to Stabilized Proteins with Native Activity

Raoul Walther, Pere Monge, Andreas Pedersen, Anja Benderoth, Jannik Pedersen, Azad Farzadfard, Ole Mandrup, Kenneth Howard, Daniel Otzen, Alexander N Zelikin*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Chemical glycosylation of proteins is a powerful tool applied widely in biomedicine and biotechnology. However, it is a challenging undertaking and typically relies on recombinant proteins and site-specific conjugations. The scope and utility of this nature-inspired methodology would be broadened tremendously by the advent of facile, scalable techniques in glycosylation, which are currently missing. In this work, we investigated a one-pot aqueous protocol to achieve indiscriminate, surface-wide glycosylation of the surface accessible amines (lysines and/or N-terminus). We reveal that this approach afforded minimal if any change in the protein activity and recognition events in biochemical and cell culture assays, but at the same time provided a significant benefit of stabilizing proteins against aggregation and fibrillation - as demonstrated on serum proteins (albumins and immunoglobulin G, IgG), an enzyme (uricase), and proteins involved in neurodegenerative disease (α-synuclein) and diabetes (insulin). Most importantly, this highly advantageous result was achieved via a one-pot aqueous protocol performed on native proteins, bypassing the use of complex chemical methodologies and recombinant proteins.

Original languageEnglish
JournalChemBioChem
Volume22
Issue14
Pages (from-to)2478-2485
Number of pages8
ISSN1439-4227
DOIs
Publication statusPublished - Jul 2021

Keywords

  • glucuronidation
  • glycosylation
  • insulin
  • protein biochemistry
  • synuclein
  • INSULIN
  • PEPTIDES
  • PRODRUGS
  • STRATEGY
  • CHEMICAL-SYNTHESIS
  • GENERATION
  • ALBUMIN

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