Portal de Periódicos da CAPES

The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans ‐splicing reaction

2009; Wiley; Volume: 583; Issue: 5 Linguagem: Inglês

10.1016/j.febslet.2009.02.003

1873-3468

J. L. Zettler, Vivien Schütz, Henning D. Mootz,

RNA modifications and cancer

We have studied the naturally split urn:x-wiley:00145793:media:feb2s001457930900101x:feb2s001457930900101x-math-si60 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 urn:x-wiley:00145793:media:feb2s001457930900101x:feb2s001457930900101x-math-si61 s −1 at 37 °C. This represents the highest rate reported for the protein trans ‐splicing reaction so far ( urn:x-wiley:00145793:media:feb2s001457930900101x:feb2s001457930900101x-math-si62 of urn:x-wiley:00145793:media:feb2s001457930900101x:feb2s001457930900101x-math-si63 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.