Congruent docking of dimeric kinesin and ncd into three-dimensional electron cryomicroscopy maps of microtubule-motor ADP complexes.
We present a new map showing dimeric kinesin bound to microtubules in the presence of ADP that was obtained by electron cryomicroscopy and image reconstruction. The directly bound monomer (first head) shows a different conformation from one in the more tightly bound empty state. This change in the first head is amplified as a movement of the second (tethered) head, which tilts upward. The atomic coordinates of kinesin.ADP dock into our map so that the tethered head associates with the bound head as in the kinesin dimer structure seen by x-ray crystallography. The new docking orientation avoids problems associated with previous predictions; it puts residues implicated by proteolysis-protection and mutagenesis studies near the microtubule but does not lead to steric interference between the coiled-coil tail and the microtubule surface. The observed conformational changes in the tightly bound states would probably bring some important residues closer to tubulin. As expected from the homology with kinesin, the atomic coordinates of nonclaret disjunctional protein (ncd).ADP dock in the same orientation into the attached head in a map of microtubules decorated with dimeric ncd.ADP. Our results support the idea that the observed direct interaction between the two heads is important at some stages of the mechanism by which kinesin moves processively along microtubules.
Hirose
K
K
National Institute of Advanced Interdisciplinary Research, Higashi, Tsukuba 305-8562, Japan.
Löwe
J
J
Alonso
M
M
Cross
R A
RA
Amos
L A
LA
D000244:Adenosine Diphosphate / Q000378:metabolism*; D001665:Binding Sites; D019281:Dimerization; D029721:Drosophila Proteins; D005615:Freezing; D007091:Image Processing, Computer-Assisted; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism*; D008854:Microscopy, Electron / Q000379:methods*; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008958:Models, Molecular; D011487:Protein Conformation; D014404:Tubulin / Q000378:metabolism
D029721:Drosophila Proteins; D014404:Tubulin; C066696:ncd protein, Drosophila; D000244:Adenosine Diphosphate; D016547:Kinesins