The SNAP-tag technology revised: an effective chemo-enzymatic approach by using a universal azide-based substrate.
J Enzyme Inhib Med Chem
; 36(1): 85-97, 2021 Dec.
Article
em En
| MEDLINE
| ID: mdl-33121288
ABSTRACT
SNAP-tag ® is a powerful technology for the labelling of protein/enzymes by using benzyl-guanine (BG) derivatives as substrates. Although commercially available or ad hoc produced, their synthesis and purification are necessary, increasing time and costs. To address this limitation, here we suggest a revision of this methodology, by performing a chemo-enzymatic approach, by using a BG-substrate containing an azide group appropriately distanced by a spacer from the benzyl ring. The SNAP-tag ® and its relative thermostable version (SsOGT-H5 ) proved to be very active on this substrate. The stability of these tags upon enzymatic reaction makes possible the exposition to the solvent of the azide-moiety linked to the catalytic cysteine, compatible for the subsequent conjugation with DBCO-derivatives by azide-alkyne Huisgen cycloaddition. Our studies propose a strengthening and an improvement in terms of biotechnological applications for this self-labelling protein-tag.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Azidas
/
Metilases de Modificação do DNA
/
Corantes Fluorescentes
Limite:
Humans
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article