Osnabrück 2002 – wissenschaftliches Programm
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MO: Molekülphysik
MO 7: Poster I (Experimentelle Techniken, Spektroskopie, Cluster)
MO 7.9: Poster
Mittwoch, 6. März 2002, 11:00–13:00, Schloss
Observation of Single Molecules during their Translocation by the Nuclear Pore Complex — •Andreas Hoekstra1, Thorsten Kues1, Daniel Rohleder1, Jan-Peter Siebrasse1, Ulrich Kubitscheck2, and Reiner Peters1 — 1Institut für Medizinische Physik und Biophysik, Robert-Koch-Str. 31, D-48149 Muenster — 2Institut für Experimentelle Physik, Postfach 330440, D-28334 Bremen
The nuclear import of karyophilic proteins occurs via the nuclear pore complex (NPC). We observed the binding of single Alexa-633 labeled secondary antibodies directed against a primary antibody decorating the filaments (NUP358) of GFP-tagged NPCs. Furthermore, single fluorescently labeled molecules of cargo import complexes were detected when attaching to and during the subsequent translocation by the NPC. For the experiments highly sensitive, high-speed laser light microscopic methods, and especially designed image processing tools were employed. As transport substrate we used fluorescently labeld P4K (500 kDa). The nuclear import was achieved using the recombinant receptor molecules karyopherin α and β, Ran and an ATP-regenerating system. Our experiments provide information on the dynamic aspects of the import process on a single molecule level.