Enhanced Delivery of Synthetic Labelled Ubiquitin into Live Cells by Using Next-Generation Ub-TAT Conjugates.
Chembiochem
; 19(24): 2553-2557, 2018 12 18.
Article
em En
| MEDLINE
| ID: mdl-30351505
ABSTRACT
Proteins and other macromolecules can be delivered into live cells by noninvasive techniques using cell-penetrating peptides. These peptides are easily synthesised by solid-phase peptide synthesis and can be conjugated onto cargo molecules to mediate cellular delivery. We designed a TAT-based cell-penetrating ubiquitin (Ub) reagent by conjugating a dimeric disulfide-linked TAT peptide to the Câ
terminus of a rhodamine-labelled Ub (RhoUb) protein. This reagent efficiently enters the cell by endocytosis and escapes from endosomes into the cytoplasm. Once the conjugate is inside the cytoplasm, the delivery vehicle is proteolytically removed by endogenous deubiquitinases (DUBs), at which point the intrinsic ubiquitination machinery is able to incorporate the RhoUb into ubiquitin conjugates. Our approach enables the controlled delivery of labelled or mutant Ub derivatives into cells, increasing our options for studying the ubiquitin system.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Rodaminas
/
Sondas Moleculares
/
Ubiquitina
/
Peptídeos Penetradores de Células
Limite:
Humans
Idioma:
En
Revista:
Chembiochem
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Holanda