The heterodimeric karyopherin functions in targeting a nuclear localization sequence (NLS)-containing protein to the nuclear pore complex followed by Ran-GTP and p10-mediated translocation of the NLS protein into the nucleoplasm. It was shown recently that Ran-GTP dissociated the karyopherin heterodimer and, in doing so, associated with karyopherin (Rexach, M., and Blobel, G.(1995) Cell 83, 683-692). We show here, using all recombinant yeast proteins expressed in Escherichia coli, that karyopherin binds to Ran-GTP and inhibits GTP hydrolysis stimulated by RanGAP (the Ran-specific GTPase activating protein). Inhibition of RanGAP-stimulated GTP hydrolysis by karyopherin was dependent on karyopherin concentration relative to Ran-GTP. Complete inhibition of RanGAP was observed at karyopherin concentrations that were equimolar to Ran-GTP. In gel filtration experiments, we found Ran-GTP and karyopherin to form a stoichiometric complex. Ran-GDP bound only weakly to karyopherin . We propose that stoichiometric complex formation between karyopherin and Ran-GTP renders Ran-GTP inaccessible to RanGAP.

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