Conformation of the Drosophila motor protein non-claret disjunctional in solution from X-ray and neutron scattering.
The quaternary structures of monomeric and dimeric Drosophila non-claret disjunctional (ncd) constructs were investigated using synchrotron x-ray and neutron solution scattering, and their low resolution shapes were restored ab initio from the scattering data. The experimental curves were further compared with those computed from crystallographic models of one monomeric and three available dimeric ncd structures in the microtubule-independent ADP-bound state. These comparisons indicate that accounting for the missing parts in the crystal structures for all these constructs is indispensable to obtain reasonable fits to the scattering patterns. A ncd construct (MC6) lacking the coiled-coil region is monomeric in solution, but the calculated scattering from the crystallographic monomer yields a poor fit to the data. A tentative configuration of the missing C-terminal residues in the form of an antiparallel beta-sheet was found that significantly improves the fit. The atomic model of a short dimeric ncd construct (MC5) without 2-fold symmetry is found to fit the data better than the symmetric models. Addition of the C-terminal residues to both head domains gives an excellent fit to the x-ray and neutron experimental data, although the orientation of the beta-sheet differs from that of the monomer. The solution structure of the long ncd construct (MC1) including complete N-terminal coiled-coil and motor domains is modeled by adding a straight coiled-coil section to the model of MC5.
Svergun
D I
DI
Institute of Crystallography, Russian Academy of Sciences, Leninsky Prospekt 59, 117333 Russia. Svergun@EMBL-Hamburg.DE
Zaccai
G
G
Malfois
M
M
Wade
R H
RH
Koch
M H
MH
Kozielski
F
F
D000818:Animals; D018360:Crystallography, X-Ray; D019281:Dimerization; D004330:Drosophila; D029721:Drosophila Proteins; D016547:Kinesins / Q000737:chemistry*; D008958:Models, Molecular; D009502:Neutrons; D011487:Protein Conformation; D017510:Protein Folding; D012996:Solutions
D029721:Drosophila Proteins; D012996:Solutions; C066696:ncd protein, Drosophila; D016547:Kinesins