Chemoselective restoration of para-azido-phenylalanine at multiple sites in proteins.
biomaterials
click-chemistry
conjugation
genetic code expansion
genomically recoded organisms
non-standard amino acids
para-azido-phenylalanine
protein engineering
Journal
Cell chemical biology
ISSN: 2451-9448
Titre abrégé: Cell Chem Biol
Pays: United States
ID NLM: 101676030
Informations de publication
Date de publication:
16 06 2022
16 06 2022
Historique:
received:
22
11
2020
revised:
02
06
2021
accepted:
30
11
2021
pubmed:
30
12
2021
medline:
22
6
2022
entrez:
29
12
2021
Statut:
ppublish
Résumé
The site-specific incorporation of nonstandard amino acids (nsAAs) during translation has expanded the chemistry and function of proteins. The nsAA para-azido-phenylalanine (pAzF) encodes a biorthogonal chemical moiety that facilitates "click" reactions to attach diverse chemical groups for protein functionalization. However, the azide moiety is unstable in physiological conditions and is reduced to para-amino-phenylalanine (pAF). Azide reduction decreases the yield of pAzF residues in proteins to 50%-60% per azide and limits protein functionalization by click reactions. Here, we describe the use of a pH-tunable diazotransfer reaction that converts pAF to pAzF at >95% efficiency in proteins. The method selectively restores pAzF at multiple sites per protein without introducing off-target modifications. This work addresses a key limitation in the production of pAzF-containing proteins by restoring azides for multi-site functionalization with diverse chemical moieties, setting the stage for the production of genetically encoded biomaterials with broad applications in biotherapeutics, materials science, and biotechnology.
Identifiants
pubmed: 34965380
pii: S2451-9456(21)00516-X
doi: 10.1016/j.chembiol.2021.12.002
pmc: PMC10173106
mid: NIHMS1891803
pii:
doi:
Substances chimiques
Amino Acids
0
Azides
0
Biocompatible Materials
0
Proteins
0
Phenylalanine
47E5O17Y3R
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1046-1052.e4Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM125951
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM141192
Pays : United States
Informations de copyright
Copyright © 2021 Elsevier Ltd. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests The authors have filed a patent related to this work. F.J.I. is a co-founder of Pearl Bio.
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