title: Regulation of ncd by the oligomeric state of tubulin.
abstract: We have compared the interaction of ncd (non-claret disjunctional), a kinesin related protein, with microtubules and tubulin heterodimer. Ultracentrifugation experiments revealed that the ncd motor domain, residues 335-700 (ncd335), does not induce tubulin polymerization but stabilizes pre-formed microtubules with a maximum effect at a 1:1 ncd335:tubulin ratio. Ncd335 binding to tubulin or microtubules was estimated by following the change in fluorescence polarization of an exogenous dye attached to Cys670 of ncd335. Ncd335 binding to tubulin (containing GTP or GDP-bound) is characterized by a 2:1 stoichiometry, a higher affinity and an increased sensitivity towards salt, ADP, ATP and AMPPNP, as compared with ncd335 binding to microtubules. Maximum ATPases were 0.06-0.08 sec(-1) and 1.8-2.0 sec(-1) for the ncd335-tubulin and ncd335-microtubules complexes, respectively. Only the polymerized complex is fully functional, suggesting the presence of additional contacts between adjacent protofilaments. Moreover, the data reveal that the oligomeric state of microtubules is a potent regulator for the activity of kinesin related proteins.
authors.affiliation: CRBM du CNRS, IFR 24, Montpellier C¿edex 05, 34293, France.
authors.forename: C
authors.initials: C
authors.lastname: Smyczynski
authors.forename: J
authors.initials: J
authors.lastname: Derancourt
authors.forename: P
authors.initials: P
authors.lastname: Chaussepied
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D001704:Biopolymers; D029721:Drosophila Proteins; D005456:Fluorescent Dyes; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009994:Osmolar Concentration; D013329:Structure-Activity Relationship; D013552:Swine; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001704:Biopolymers; D029721:Drosophila Proteins; D005456:Fluorescent Dyes; D014404:Tubulin; C066696:ncd protein, Drosophila; D000251:Adenosine Triphosphatases; D016547:Kinesins