Solution structures of dimeric kinesin and ncd motors.
The dimeric structure of the members of the kinesin family of motor proteins determines the individual characteristics of their microtubule-based motility. Crystal structures for ncd and kinesin dimers, which move in opposite directions on microtubules, show possible states of these dimers with ADP bound but give no information about these dimers in solution. Here, low-angle X-ray and neutron scattering were used to investigate their solution structures. Scattering profiles of Drosophila ncd 281-700 (NCD281) and human kinesin 1-420 (hKIN420) were compared with models made from the crystallographically determined structures of NCD281 and rat kinesin 1-379 (rKIN379). From the low-angle region it was found that the radius of gyration (Rg) of NCD281 is 3.60 +/- 0.075 nm, which is in agreement with the crystallography-based model. Scattering by longer ncd constructs (NCD250 and NCD224) is also well fit by the appropriate crystallography-based models. However, the measured Rg of hKIN420, 4.05 +/- 0.075 nm, is significantly smaller than that of the crystallography-based model. In addition, the overall scattering pattern of NCD281 is well fit by the model, but that of hKIN420 is poorly fit. Model calculations indicate that the orientation of the catalytic cores is different from that observed in the rKIN379 crystal structure. Like the crystal structure, the best-fitting models do not show 2-fold symmetry about the neck axis; however, their overall shape more resembles a mushroom than the "T"-like orientation of the catalytic cores found in the crystal structure. The center of mass separations of the catalytic cores in the best-fitting models are 0.7-1 nm smaller than in the crystal structure.
Stone
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DB
Cardiovascular Research Institute, Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0130, USA.
Hjelm
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Mendelson
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RA
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000818:Animals; D002384:Catalysis; D018360:Crystallography, X-Ray; D019281:Dimerization; D004330:Drosophila; D029721:Drosophila Proteins; D006801:Humans; D016547:Kinesins / Q000737:chemistry*; D008958:Models, Molecular; D009502:Neutrons; D017433:Protein Structure, Secondary; D051381:Rats; D012542:Scattering, Radiation; D012996:Solutions; D014965:X-Rays
D029721:Drosophila Proteins; D012996:Solutions; C066696:ncd protein, Drosophila; D000251:Adenosine Triphosphatases; D016547:Kinesins