Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein.
Acta Biochim Biophys Sin (Shanghai)
; 55(8): 1310-1318, 2023 Jul 23.
Article
em En
| MEDLINE
| ID: mdl-37489009
ABSTRACT
Intein-mediated protein splicing has been widely used in protein engineering; however, the splicing efficiency and extein specificity usually limit its further application. Thus, there is a demand for more general inteins that can overcome these limitations. Here, we study the trans-splicing of CPE intein obtained from the directed evolution of Cne PRP8, which shows that its splicing rate is ~29- fold higher than that of the wild-type. When the +1 residue of C-extein is changed to cysteine, CPE also shows high splicing activity. Faster association and higher affinity may contribute to the high splicing rate compared with wild-type intein. These findings have important implications for the future engineering of inteins and provide clues for fundamental studies of protein structure and folding.
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Base de dados:
MEDLINE
Assunto principal:
Processamento de Proteína
/
Cryptococcus neoformans
/
Inteínas
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article