Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein Directed evolution of the

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...

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Bibliographic Details
Published inActa biochimica et biophysica Sinica Vol. 55; no. 8; pp. 1310 - 1318
Main Authors Zhan, Qin, Shi, Changhua, Jiang, Yu, Gao, Xianling, Lin, Ying
Format Journal Article
LanguageEnglish
Published Oxford University Press 23.07.2023
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Summary: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.
Bibliography:These authors contributed equally to this work.
ISSN:1672-9145
1745-7270
DOI:10.3724/abbs.2023135