Generation and Characterization of Engineered Ubiquitin Variants to Modulate the Ubiquitin Signaling Cascade.
Cold Spring Harb Protoc
; 2024(4): pdb.over107784, 2024 Apr 01.
Article
em En
| MEDLINE
| ID: mdl-36997275
ABSTRACT
The ubiquitin signaling cascade plays a crucial role in human cells. Consistent with this, malfunction of ubiquitination and deubiquitination is implicated in the initiation and progression of numerous human diseases, including cancer. Therefore, the development of potent and specific modulators of ubiquitin signal transduction has been at the forefront of drug development. In the past decade, a structure-based combinatorial protein-engineering approach has been used to generate ubiquitin variants (UbVs) as protein-based modulators of multiple components in the ubiquitin-proteasome system. Here, we review the design and generation of phage-displayed UbV libraries, including the processes of binder selection and library improvement. We also provide a comprehensive overview of the general in vitro and cellular methodologies involved in characterizing UbV binders. Finally, we describe two recent applications of UbVs for developing molecules with therapeutic potential.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Ubiquitina
/
Técnicas de Visualização da Superfície Celular
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article