KPNA7 contains an N-terminal IBB. (A) KPNA proteins were translated as 35S-methionine labeled polypeptides and used in binding assays with GST-Importin β. (B) Full-length and ΔIBB forms of KPNA7 tested for binding to Importin β. (C) Importin β binding to the IBB of KPNA7 is RanGTP-sensitive. Recombinant Importin β binding to GST-IBB proteins was performed in the absence and presence of recombinant RanGTP, and by subsequent addition of RanGTP to preformed GST-IBB-importin β complexes. These assays were performed with the GTPase deficient RanQ69L mutant. Bound and eluted fractions were examined by immunoblotting using a monoclonal antibody to Importin β. (D) KPNA1, KPNA2, KPNA7, KPNA7 ΔIBB binding to GST-SV40 NLS using 35S-methionine labeled polypeptides.
Kelley et al. BMC Cell Biology 2010 11:63 doi:10.1186/1471-2121-11-63