A new preparation method of covalent annular nanodiscs based on MTGase.
Arch Biochem Biophys
; 756: 109997, 2024 06.
Article
en En
| MEDLINE
| ID: mdl-38621443
ABSTRACT
The preservation of the native conformation and functionality of membrane proteins has posed considerable challenges. While detergents and liposome reconstitution have been traditional approaches, nanodiscs (NDs) offer a promising solution by embedding membrane proteins in phospholipids encircled by an amphipathic helical protein MSP belt. Nevertheless, a drawback of commonly used NDs is their limited homogeneity and stability. In this study, we present a novel approach to construct covalent annular nanodiscs (cNDs) by leveraging microbial transglutaminase (MTGase) to catalyze isopeptide bond formation between the side chains of terminal amino acids, specifically Lysine (K) and Glutamine (Q). This methodology significantly enhances the homogeneity and stability of NDs. Characterization of cNDs and the assembly of membrane proteins within them validate the successful reconstitution of membrane proteins with improved homogeneity and stability. Our findings suggest that cNDs represent a more suitable tool for investigating interactions between membrane proteins and lipids, as well as for analyzing membrane protein structures.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Transglutaminasas
/
Nanoestructuras
/
Proteínas de la Membrana
Idioma:
En
Revista:
Arch Biochem Biophys
/
Arch. biochem. biophys
/
Archives of biochemistry and biophysics
Año:
2024
Tipo del documento:
Article
País de afiliación:
China