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. 2006 Sep 25;174(7):1035–1045. doi: 10.1083/jcb.200606003

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Copyright © 2006, The Rockefeller University Press

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Figure 1. — Preparation and characterization of recombinant C. elegans kinesin-II and OSM-3. (A) SDS gels of Sf9 cell high speed supernatant (left), Talon column eluate (middle), and Sephacryl S-300 purified kinesin-II (right). (B) SDS gels of purified kinesin-II and OSM-3. (C and D) Double reciprocal plots of kinesin-II– (C) and OSM-3 (D)–driven MT motility versus [Mg-ATP] in standard MT gliding assays. (E) MT gliding velocity driven by kinesin-II (circles), OSM-3 (squares), and OSM-3–G444E (triangles) under standard assay conditions but varying concentrations of K₂-Pipes. Error bars represent the standard deviations. (F and G) On sucrose density gradients (F) and gel filtration columns (G), the KLP-11, KAP-1, and KLP-20 subunits elute as a monodisperse heterotrimeric complex (S value = 9.8; Rs = 7.1 nm; and native molecular mass = 287 kD) in a KLP-11/KLP-20/KAP-1 molar stoichiometry of 1.0:1.17:0.89 (protein standard peak positions are also indicated).