Highly efficient protein trans-splicing by a naturally split DnaE intein from Nostoc punctiforme

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• 作者：Hideo Iwai ; Sara Zü ; ger ; Jennifer Jin and Pui-Hang Tam
• 关键词：Protein splicing ; Intein ; Segmental isotopic labelling ; NMR
• 刊名：FEBS Letters
• 出版年：2006
• 期刊代码：70_00145793
• 类别：bio
• 出版时间：20 March 2006
• 卷：580
• 期：7
• 页码：1853-1858
• 文件大小：380 K

摘要

Protein trans-splicing by the naturally split intein of the gene dnaE from Nostoc punctiforme (Npu DnaE) was demonstrated here with non-native exteins in Escherichia coli. Npu DnaE possesses robust trans-splicing activity with an efficiency of >98%, which is superior to that of the DnaE intein from Synechocystis sp. strain PCC6803 (Ssp DnaE). Both the N- and C-terminal parts of the split Npu DnaE intein can be substituted with the corresponding fragment of Ssp DnaE without loss of trans-splicing activity. Protein splicing with the Npu DnaE_N is also more tolerant of amino acid substitutions in the C-terminal extein sequence.