Improving Split Reporters of Protein-Protein Interactions through Orthology-Based Protein Engineering.
ACS Chem Biol
; 19(2): 428-441, 2024 02 16.
Article
em En
| MEDLINE
| ID: mdl-38289242
ABSTRACT
Protein-protein interactions (PPIs) can be detected through selective complementation of split fluorescent reporters made of two complementary fragments that reassemble into a functional fluorescent reporter when in close proximity. We previously introduced splitFAST, a chemogenetic PPI reporter with rapid and reversible complementation. Here, we present the engineering of splitFAST2, an improved reporter displaying higher brightness, lower self-complementation, and higher dynamic range for optimal monitoring of PPI using an original protein engineering strategy that exploits proteins with orthology relationships. Our study allowed the identification of a system with improved properties and enabled a better understanding of the molecular features controlling the complementation properties. Because of the rapidity and reversibility of its complementation, its low self-complementation, high dynamic range, and improved brightness, splitFAST2 is well suited to study PPI with high spatial and temporal resolution, opening great prospects to decipher the role of PPI in various biological contexts.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Mapeamento de Interação de Proteínas
Idioma:
En
Revista:
ACS Chem Biol
Ano de publicação:
2024
Tipo de documento:
Article