Toward understanding the structure and interactions of microtubules and motor proteins.
To obtain an overall three-dimensional picture of the interaction between microtubules and the motor proteins of the kinesin family it will be necessary to take account of both atomic resolution structures obtained by X-ray crystallography and medium resolution reconstructions obtained by electron cryomicroscopy. We examine the problems associated with obtaining the required structural information from electron micrographs of vitreous ice-embedded microtubules decorated with motor domains. We find that the minus-end directed motor, ncd, decorates microtubules with an 80 A periodicity as for kinesin. Our theoretical analysis and experiments with ncd illustrate the difficulty in determining unambiguously the surface lattice organization by diffraction analysis of micrographs. 3D reconstructions of decorated microtubules are required to accurately locate the motor domains. Helical diffraction theory is not usually applicable because microtubules are cylindrical structures that rarely have complete helical symmetry. We propose using a back-projection method based on the long pitch helices formed by individual protofilaments. Model reconstructions show that this approach is feasible.
Wade
R H
RH
Laboratoire de Microscopie Electronique Structurale, Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France.
Horowitz
R
R
Milligan
R A
RA
D003198:Computer Simulation; D018360:Crystallography, X-Ray / Q000379:methods; D016547:Kinesins / Q000737:chemistry / Q000378:metabolism; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D008870:Microtubules / Q000648:ultrastructure*; D008961:Models, Structural; D011487:Protein Conformation; D017433:Protein Structure, Secondary
D008869:Microtubule-Associated Proteins; D016547:Kinesins