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The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans ‐splicing reaction
Author(s) -
Zettler Joachim,
Schütz Vivien,
Mootz Henning D.
Publication year - 2009
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2009.02.003
Subject(s) - intein , protein splicing , rna splicing , protein subunit , chemistry , biochemistry , biology , gene , rna
We have studied the naturally splitαsubunit 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 ) × 10 ‐ 2s −1 at 37 °C. This represents the highest rate reported for the protein trans ‐splicing reaction so far (t 1 / 2of∼60 s). Furthermore, the reaction was very robust and high‐yielding with respect to different extein sequences, temperatures from 6 to 37 °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.