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

doi:10.1002/cbic.202100228

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 af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

1 Citationer (Scopus)

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.

Originalsprog	Engelsk
Tidsskrift	ChemBioChem
Vol/bind	22
Nummer	14
Sider (fra-til)	2478-2485
Antal sider	8
ISSN	1439-4227
DOI	https://doi.org/10.1002/cbic.202100228
Status	Udgivet - jul. 2021

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10.1002/cbic.202100228

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

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.",

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author = "Raoul Walther and Pere Monge and Andreas Pedersen and Anja Benderoth and Jannik Pedersen and Azad Farzadfard and Ole Mandrup and Kenneth Howard and Daniel Otzen and Zelikin, {Alexander N}",

note = "{\textcopyright} 2021 Wiley-VCH GmbH.",

year = "2021",

month = jul,

doi = "10.1002/cbic.202100228",

language = "English",

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TY - JOUR

T1 - Per-glycosylation of the Surface-Accessible Lysines

T2 - One-Pot Aqueous Route to Stabilized Proteins with Native Activity

AU - Walther, Raoul

AU - Monge, Pere

AU - Pedersen, Andreas

AU - Benderoth, Anja

AU - Pedersen, Jannik

AU - Farzadfard, Azad

AU - Mandrup, Ole

AU - Howard, Kenneth

AU - Otzen, Daniel

AU - Zelikin, Alexander N

PY - 2021/7

Y1 - 2021/7

N2 - 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.

AB - 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.

KW - glucuronidation

KW - glycosylation

KW - insulin

KW - protein biochemistry

KW - synuclein

KW - INSULIN

KW - PEPTIDES

KW - PRODRUGS

KW - STRATEGY

KW - CHEMICAL-SYNTHESIS

KW - GENERATION

KW - ALBUMIN

U2 - 10.1002/cbic.202100228

DO - 10.1002/cbic.202100228

M3 - Journal article

C2 - 33998129

SN - 1439-4227

VL - 22

SP - 2478

EP - 2485

JO - ChemBioChem

JF - ChemBioChem

IS - 14

ER -

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

Abstract

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