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 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 DnaEN is also more tolerant of amino acid substitutions in the C-terminal extein sequence.
Iwai
Hideo
H
Department of Chemistry, 110 Science Place, Saskatoon, SK, Canada, S7N 5C9. hideo.iwai@helsinki.fi
Züger
Sara
S
Jin
Jennifer
J
Tam
Pui-Hang
PH
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis*; D004258:DNA Polymerase III / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000502:physiology; D047668:Inteins / Q000502:physiology*; D046937:Nostoc / Q000201:enzymology*; D010446:Peptide Fragments; D019154:Protein Splicing; D046939:Synechocystis / Q000201:enzymology
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D010446:Peptide Fragments; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III