The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction.
We have studied the naturally split alpha subunit of the DNA polymerase III (DnaE) intein from Nostoc punctiforme PCC73102 (Npu) using purified proteins and determined an apparent first-order rate constant of (1.1+/-0.2)x10(-2) s(-1) at 37 degrees C. This represents the highest rate reported for the protein trans-splicing reaction so far (t(1/2) of approximately 60s). Furthermore, the reaction was very robust and high-yielding with respect to different extein sequences, temperatures from 6 to 37 degrees C, and the presence of up to 6 M urea. Given these outstanding properties, the Npu DnaE intein appears to be the intein of choice for many applications in protein and cellular chemistry.
Zettler
Joachim
J
Technische Universität Dortmund, Fakultät Chemie-Chemische Biologie, Dortmund, Germany.
Schütz
Vivien
V
Mootz
Henning D
HD
D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D047668:Inteins; D046937:Nostoc / Q000235:genetics* / Q000378:metabolism*; D017354:Point Mutation / Q000235:genetics; D013997:Time Factors; D020040:Trans-Splicing / Q000235:genetics*
C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III