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Peptide Synthesis> | The fluorescein-labeled pol NLS peptide: S3KRKAPQETLNGG14-Lys(FITC), used for fluorescence polarization assays, was obtained from Genscript at a purity level of > 90% C-terminal Histagged human Importin 5, corresponding to residues 66–512, (hImp 5IBB) was purchased from Genscript and cloned into the pET30 expression vector | Get A Quote |
DNA polymerase (pol ) requires nuclear localization to fulfil its DNA repair function. Although its small size has been interpreted to imply the absence of a need for active nuclear import, sequence and structural analysis suggests that a monopartite nuclear localization signal (NLS) may reside in the Nterminal lyase domain. Binding of this domain to Importin 1 (Imp1) was confirmed by gel filtration and NMR studies. Affinity was quantified by fluorescence polarization analysis of a fluorescein-tagged peptide corresponding to pol residues 2–13. These studies indicate high affinity binding, characterized by a low micromolar Kd, that is selective for the murine Importin 1 (mImp1) minor site, with the Kd strength... More