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

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 f...

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Bibliographic Details
Published in:FEBS letters 2006-03, Vol.580 (7), p.1853-1858
Main Authors: Iwai, Hideo, Züger, Sara, Jin, Jennifer, Tam, Pui-Hang
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacterial Proteins - biosynthesis
DNA Polymerase III - physiology
Escherichia coli
Escherichia coli Proteins - physiology
GB1
Intein
Inteins - physiology
IPTG
isopropyl β-d-thiogalactoside
MALDI-TOF
matrix-assisted laser desorption and ionization-time-of-flight
NMR
Nostoc - enzymology
Nostoc punctiforme
Peptide Fragments
Protein Splicing
SDS–PAGE
Segmental isotopic labelling
sodium dodecyl sulfate–polyacrylamide gel electrophoresis
Synechocystis
Synechocystis - enzymology
the B1 domain of IgG binding protein, protein G
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Summary: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.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2006.02.045