title: Recombinant kinesin motor domain binds to beta-tubulin and decorates microtubules with a B surface lattice.
abstract: We have expressed the recombinant squid kinesin head domain in Escherichia coli and studied its interaction with microtubules. The head is active as a microtubule-stimulated ATPase and binds to microtubules, but it does not support microtubule gliding by itself. The head binds to both microtubules and depolymerized tubulin. In each case the zero-length crosslinker 1-ethyl-3-[3-dimethylamino)propyl] carbodiimide induces a bond specifically to beta- but not alpha-tubulin. The head decorates brain microtubules with an 8-nm axial spacing. Thus the stoichiometry is one kinesin head per tubulin dimer. The lattice is that of flagellar B-tubules, implying that reassembled microtubules are not symmetric. Moreover, the A- and B-tubules of intact flagellar outer doublets are both decorated with a B lattice. This suggests that the B lattice is a general property of microtubules.
authors.affiliation: Max Planck Unit for Structural Molecular Biology, Deutsches Elektronen Synchrotron, Hamburg, Germany.
authors.forename: Y H
authors.initials: YH
authors.lastname: Song
authors.forename: E
authors.initials: E
authors.lastname: Mandelkow
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D003432:Cross-Linking Reagents; D049832:Decapodiformes; D005407:Flagella / Q000648:ultrastructure; D066298:In Vitro Techniques; D016547:Kinesins / Q000378:metabolism*; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000201:enzymology* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D012617:Sea Urchins / Q000648:ultrastructure; D014404:Tubulin / Q000378:metabolism*
chemical_list: D003432:Cross-Linking Reagents; D008869:Microtubule-Associated Proteins; D011994:Recombinant Proteins; D014404:Tubulin; D000251:Adenosine Triphosphatases; D016547:Kinesins