Three-dimensional cryoelectron microscopy of dimeric kinesin and ncd motor domains on microtubules.
Kinesin and ncd motor proteins are homologous in sequence yet move in opposite directions along microtubules. We have previously shown that monomeric kinesin and ncd bind in the same orientation on equivalent sites relative to the ends of tubulin sheets of known polarity. We now report cryoelectron microscope images of 16-protofilament microtubules decorated with both single- and double-headed kinesin and double-headed ncd. Three-dimensional density maps and difference maps show that, in adenosine 5'-[beta,gamma-imido]triphosphate, both dimeric motors bind tightly to microtubules via one head, leaving the other free, though apparently in a fixed position. The attached heads of dimers bind to tubulin in the same way as single kinesin heads. The second heads are connected to the tops of the first but, whereas the second kinesin head is closely associated with the first, pairs of ncd heads are splayed apart. There is also a distinct difference in orientation: the second kinesin head is tilted toward the microtubule plus end, while the second head of ncd points toward the minus end.
Hirose
K
K
Medical Research Council Laboratory of Molecular Biology, Cambridge, United Kingdom.
Lockhart
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A
Cross
R A
RA
Amos
L A
LA
D000244:Adenosine Diphosphate; D000266:Adenylyl Imidodiphosphate; D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D029721:Drosophila Proteins; D006135:Gryllidae; D007091:Image Processing, Computer-Assisted; D016547:Kinesins / Q000737:chemistry / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D008868:Microtubule Proteins / Q000737:chemistry / Q000648:ultrastructure*; D008870:Microtubules / Q000737:chemistry / Q000648:ultrastructure*; D011487:Protein Conformation; D051381:Rats; D013737:Testis / Q000737:chemistry
D029721:Drosophila Proteins; D008868:Microtubule Proteins; C066696:ncd protein, Drosophila; D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D016547:Kinesins