Proximity-Based Sortase-Mediated Ligation.
Angew Chem Int Ed Engl
; 56(19): 5349-5352, 2017 05 02.
Article
en En
| MEDLINE
| ID: mdl-28374553
ABSTRACT
Protein bioconjugation has been a crucial tool for studying biological processes and developing therapeutics. Sortaseâ
A (SrtA), a bacterial transpeptidase, has become widely used for its ability to site-specifically label proteins with diverse functional moieties, but a significant limitation is its poor reaction kinetics. In this work, we address this by developing proximity-based sortase-mediated ligation (PBSL), which improves the ligation efficiency to over 95 % by linking the target protein to SrtA using the SpyTag-SpyCatcher peptide-protein pair. By expressing the target protein with SpyTag C-terminal to the SrtA recognition motif, it can be covalently captured by an immobilized SpyCatcher-SrtA fusion protein during purification. Following the ligation reaction, SpyTag is cleaved off, rendering PBSL traceless, and only the labeled protein is released, simplifying target protein purification and labeling to a single step.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Péptidos
/
Proteínas Bacterianas
/
Cisteína Endopeptidasas
/
Aminoaciltransferasas
Idioma:
En
Revista:
Angew Chem Int Ed Engl
Año:
2017
Tipo del documento:
Article
País de afiliación:
Estados Unidos