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. Author manuscript; available in PMC: 2009 Jun 20.
Published in final edited form as: Mol Cell. 2008 Jun 20;30(6):721–731. doi: 10.1016/j.molcel.2008.04.022

Figure 1. Nup107 and Nup133 Interact in Tail-to-Tail Fashion.

Figure 1

(A) In vitro binding of full length human Nup133 to recombinant GST-tagged human Nup107 visualized by autoradiography. Top and bottom panels show the bound and unbound fractions of [35S] methionine-labeled Nup133 translation product incubated with recombinant GST-Nup107 proteins immobilized on affinity resin. The C-terminus of Nup107 is critical for its interaction with Nup133. (B) Various C-terminal fragments of Nup133 (marked with asterisks) were co-expressed with GST-tagged Nup107 (aa 658–925) from separate vectors in E. coli and co-purified by glutathione sepharose affinity chromatography. Total cell lysates (t) and elutions from the GST-affinity columns (e) were separated by SDS-PAGE and analyzed by Coomassie staining. (C) Schematic representation of Nup133 constructs tested for Nup107-binding (+). A construct spanning residues 934 to 1156 of Nup133 represents the minimum domain required for the interaction with Nup107. The relative amounts of co-expressed proteins differs from clone to clone, due to pGEX- and pET-derived vectors harboring the same origin of replication. The results of the binding experiment are therefore limited to qualitative interpretation.