Authors: Thomas Zacharchenko Igor Barsukov Daniel J Rigden Daimark Bennett Olga Mayans
Publish Date: 2016/09/03
Volume: 35, Issue: 5, Pages: 340-345
Abstract
Human protein phosphatase 1 nuclear targeting subunit PNUTS plays critical roles in DNA repair cell growth and survival The Nterminal domain of PNUTS mediates interactions with Tox4 and the phosphatase and tensin homolog PTEN which are essential for the roles of this protein To study this Nterminal domain we have established its recombinant overproduction in E coli utilizing NusA fusion Upon removal of the tag the remaining PNUTS sample is soluble and highly pure We have characterized the domain using circular dichroism and nuclear magnetic resonance and analyzed its sequence using bioinformatics All data agree in suggesting that the PNUTS Nterminal segment adopts a compact globular fold rich in αhelical content where the folded fraction is substantially larger than the previously annotated fold We conclude that this domain adopts a single fold likely being an extended form of the transcription factor SII leucine/tryptophan conservedmotif Thermal denaturation yielded a melting temperature of ~495 °C confirming the stability of the fold These findings pave the way for the molecular characterization of functional interactions mediated by the Nterminal region of PNUTS
Keywords: