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

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 -splicing of CPE intei...

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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 China China Science Publishing & Media Ltd 01.08.2023
Subjects
C-extein
Cne PRP8 intein
directed evolution
C-extein
PRP8 intein
directed evolution
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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 -splicing of CPE intein obtained from the directed evolution of PRP8, which shows that its splicing rate is ~29- 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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ISSN:1672-9145
1745-7270
DOI:10.3724/abbs.2023135