X-ray Structure and Microtubule Interaction of the Motor Domain of Neurospora crassa NcKin3, a Kinesin with Unusual Processivity(,).

@article{citeulike:2277410, abstract = {Neurospora crassa kinesin NcKin3 belongs to a unique fungal-specific subgroup of small Kinesin-3-related motor proteins. One of its functions appears to be the transport of mitochondria along microtubules. Here, we present the X-ray structure of a C-terminally truncated monomeric construct of NcKin3 comprising the motor domain and the neck linker, and a 3-D image reconstruction of this motor domain bound to microtubules, by cryoelectron microscopy. The protein contains Mg.ADP bound to the active site, yet the structure resembles an ATP-bound state. By comparison with structures of the Kinesin-3 motor Kif1A in different nucleotide states (Kikkawa, M. et al. (2001) Nature (London, U.K.) 411, 439-445), the NcKin3 structure corresponds to the AMPPCP complex of Kif1A rather than the AMPPNP complex. NcKin3-specific differences in the coordination of the nucleotide and asymmetric interactions between adjacent molecules in the crystal are discussed in the context of the unusual kinetics of the dimeric wild-type motor and the monomeric construct used for crystal structure analysis. The NcKin3 motor decorates microtubules at a stoichiometry of one head per alphabeta-tubulin heterodimer, thereby forming an axial periodicity of 8 nm. In spite of unusual extensions at the N-terminus and within flexible loops L2, L8a, and L12 (corresponding to the K-loop of monomeric kinesins), the microtubule binding geometry is similar to that of other members of the kinesin family.}, address = {Max-Planck-Unit for Structural Molecular Biology, Notkestrasse 85, 22607 Hamburg, Germany, Institute for Cell Biology, Ludwig-Maximilians-University Munich, Schillerstrasse 42, 80336, Germany, and Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347.}, author = {Marx, Alexander and M\"{u}ller, Jens and Mandelkow, Eva-Maria M. and Woehlke, G\"{u}nther and Bouchet-Marquis, Cedric and Hoenger, Andreas and Mandelkow, Eckhard }, citeulike-article-id = {2277410}, doi = {10.1021/bi701483h}, issn = {0006-2960}, journal = {Biochemistry}, keywords = {kinesin, motor, xray-crystal-structure}, month = {January}, posted-at = {2008-02-29 10:59:03}, priority = {2}, title = {X-ray Structure and Microtubule Interaction of the Motor Domain of Neurospora crassa NcKin3, a Kinesin with Unusual Processivity(,).}, url = {http://dx.doi.org/10.1021/bi701483h}, year = {2008} }

TY - JOUR ID - citeulike:2277410 L3 - citeulike-article-id:2277410 TI - X-ray Structure and Microtubule Interaction of the Motor Domain of Neurospora crassa NcKin3, a Kinesin with Unusual Processivity(,). JF - Biochemistry AD - Max-Planck-Unit for Structural Molecular Biology, Notkestrasse 85, 22607 Hamburg, Germany, Institute for Cell Biology, Ludwig-Maximilians-University Munich, Schillerstrasse 42, 80336, Germany, and Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309-0347. SN - 0006-2960 N2 - Neurospora crassa kinesin NcKin3 belongs to a unique fungal-specific subgroup of small Kinesin-3-related motor proteins. One of its functions appears to be the transport of mitochondria along microtubules. Here, we present the X-ray structure of a C-terminally truncated monomeric construct of NcKin3 comprising the motor domain and the neck linker, and a 3-D image reconstruction of this motor domain bound to microtubules, by cryoelectron microscopy. The protein contains Mg.ADP bound to the active site, yet the structure resembles an ATP-bound state. By comparison with structures of the Kinesin-3 motor Kif1A in different nucleotide states (Kikkawa, M. et al. (2001) Nature (London, U.K.) 411, 439-445), the NcKin3 structure corresponds to the AMPPCP complex of Kif1A rather than the AMPPNP complex. NcKin3-specific differences in the coordination of the nucleotide and asymmetric interactions between adjacent molecules in the crystal are discussed in the context of the unusual kinetics of the dimeric wild-type motor and the monomeric construct used for crystal structure analysis. The NcKin3 motor decorates microtubules at a stoichiometry of one head per alphabeta-tubulin heterodimer, thereby forming an axial periodicity of 8 nm. In spite of unusual extensions at the N-terminus and within flexible loops L2, L8a, and L12 (corresponding to the K-loop of monomeric kinesins), the microtubule binding geometry is similar to that of other members of the kinesin family. KW - kinesin KW - motor KW - xray-crystal-structure AU - Marx, Alexander AU - Müller, Jens AU - Mandelkow, Eva-Maria AU - Woehlke, Günther AU - Bouchet-Marquis, Cedric AU - Hoenger, Andreas AU - Mandelkow, Eckhard PY - 2008/01/19/ UR - http://dx.doi.org/10.1021/bi701483h ER -