Effective production of oligomeric membrane proteins by EarlyBac-insect cell system.
Methods Enzymol
; 653: 3-19, 2021.
Article
en En
| MEDLINE
| ID: mdl-34099177
ABSTRACT
Despite major advances in methodologies for membrane protein production over the last two decades, there remain challenging protein complexes that are technically difficult to yield by conventional recombinant expression methods. A large number of these proteins are multimeric membrane proteins from eukaryotic species, which are required to pass through stringent quality control mechanisms of host cells for proper folding and complex assembly. Here, we describe the development procedure to improve the production efficiency of multi-oligomeric membrane protein complexes in insect cells and recombinant baculovirus, which involves screening of promoters, enhancers, and untranslated regions for expression levels, using calcium homeostasis modulator (CALHM) and N-methyl-d-aspartate receptor (NMDAR) proteins as examples. We demonstrate that our insect cell expression strategy is effective in expression of both multi-homomeric CALHM proteins and multi-heteromeric NMDARs.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Baculoviridae
/
Proteínas de la Membrana
Límite:
Animals
Idioma:
En
Revista:
Methods Enzymol
Año:
2021
Tipo del documento:
Article