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X-ray crystal structure of the yeast KAR3 motor domain complexed with MgADP to 2.3 Å resolution
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The X-ray structure of the head domain of the minus-end directed motor Kar3 from yeast shows similarities with the motor domains of Ncd and kinesin. This similarity between different core motor domains implies that the directionality is specified by elements outside the core.
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The first structure of tubulin at near-atomic detail was solved by electron crystallography at a resolution of 3.7 Å. This work revealed the structure of the αβ-tubulin heterodimer in the presence of taxol. It showed the positions of the nucleotides and taxol and revealed directly the structure of the microtubule protofilament, resolving both the inter-dimer as well as the intra-dimer contacts.
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Nogales E, Wolf S, Downing K Structure of the αβ tubulin dimer by electron crystallography. Nature. 391:1998;199-203. The first structure of tubulin at near-atomic detail was solved by electron crystallography at a resolution of 3.7 Å. This work revealed the structure of the αβ-tubulin heterodimer in the presence of taxol. It showed the positions of the nucleotides and taxol and revealed directly the structure of the microtubule protofilament, resolving both the inter-dimer as well as the intra-dimer contacts.
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••], was fitted into the map of microtubules obtained by helical image reconstruction. The model shows the axial and lateral contacts between tubulin subunits in a microtubule
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••], was fitted into the map of microtubules obtained by helical image reconstruction. The model shows the axial and lateral contacts between tubulin subunits in a microtubule.
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The interaction of kinesin with microtubules has been investigated by screening the kinesin surface with alanine mutations. The authors proposed the region around the helices (4-α5 and loop 11) as the predominant microtubule interaction zone.
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Proteolytic mapping of kinesin/Ncd-microtubule interface: Nucleotide-dependent conformational-changes in the loops L8 and L12
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The authors used an array of proteases to investigate the footprint of ncd and kinesin by comparing their cleavage in the microtubule-bound and unbound states. Microtubule binding protects an area between loops 8, 11 and 12, in agreement with results from site-directed mutagenesis.
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A study of chimeras between ncd and kinesin to investigate the origin of directionality within these two oppositely directed molecular motors. The linker and neck regions of these molecules were identified as key elements of directional walking. Depending on length and position of the exchanged linker/neck regions they found that the directionality of these motors even to be reversed.
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Case RB, Pierce DW, Hom-Booher N, Hart CL, Vale RD The directional preference of kinesin motors is specified by an element outside of the motor catalytic domain. Cell. 90:1997;959-966. A study of chimeras between ncd and kinesin to investigate the origin of directionality within these two oppositely directed molecular motors. The linker and neck regions of these molecules were identified as key elements of directional walking. Depending on length and position of the exchanged linker/neck regions they found that the directionality of these motors even to be reversed.
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Case, R.B.1
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Reversal of the direction of movement of a molecular motor
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••] this paper demonstrates the role of the kinesin linker region for their directionality
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Using ncd-kinesin chimeras the authors studied the role of linker/neck region for residues important for directionality. Specific mutations close to the core motor domain were found to restore the reversed directionality of the chimeras to the original one.
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Endow SA, Waligora KW Determinants of kinesin motor polarity. Science. 281:1998;1200-1202. Using ncd-kinesin chimeras the authors studied the role of linker/neck region for residues important for directionality. Specific mutations close to the core motor domain were found to restore the reversed directionality of the chimeras to the original one.
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Demonstration of coiled-coil interactions within the kinesin neck region using synthetic peptides - implications for motor activity
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••]. They also identified several 'non-ideal' residues within the two central heptads, suggesting a destabilization of that sequence.
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••]. They also identified several 'non-ideal' residues within the two central heptads, suggesting a destabilization of that sequence.
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Identification of kinesin neck region as a stable alpha-helical coiled coil and its thermodynamic characterization
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Morii H, Takenawa T, Arisaka F, Shimizu T Identification of kinesin neck region as a stable alpha-helical coiled coil and its thermodynamic characterization. Biochemistry. 36:1997;1933-1942.
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Interaction of monomeric and dimeric kinesin with microtubules
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Determination of the kinesin-microtubule binding stoichiometry by scanning transmission electron microscopy, and demonstration of a 16 nm repeat on the decorated microtubule lattice which has important implications for kinesin binding and motility.
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Thormahlen M, Marx A, Muller SA, Song Y-H, Mandelkow E-M, Aebi U, Mandelkow E Interaction of monomeric and dimeric kinesin with microtubules. J Mol Biol. 275:1998;795-809. Determination of the kinesin-microtubule binding stoichiometry by scanning transmission electron microscopy, and demonstration of a 16 nm repeat on the decorated microtubule lattice which has important implications for kinesin binding and motility.
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