The shapes of the motor domains of two oppositely directed microtubule motors, ncd and kinesin: a neutron scattering study.
The shapes of the motor domains of kinesin and ncd, which move in opposite directions along microtubules, have been investigated. Using proteins expressed in Escherichia coli, it was found that at high salt (> 200 mM) Drosophila ncd motor domain (R335-K700) and human kinesin motor domain (M1-E349) were both sufficiently monomeric to allow an accurate determination of their radii of gyration (Rg) and their molecular weights. The measured Rg values of the ncd and kinesin motor domains in D2O were 2.06 +/- 0.06 and 2.05 +/- 0.04 nm, respectively, and the molecular weights were consistent with those computed from the amino acid compositions. Fitting of the scattering curves to approximately 3.5 nm resolution showed that the ncd and kinesin motor domains can be described adequately by triaxial ellipsoids having half-axes of 1.42 +/- 0.38, 2.24 +/- 0.44, and 3.65 +/- 0.22 nm, and half-axes of 1.52 +/- 0.23, 2.00 +/- 0.25, and 3.73 +/- 0.10 nm, respectively. Both motor domains are described adequately as somewhat flattened prolate ellipsoids with a maximum dimension of approximately 7.5 nm. Thus, it appears that the overall shapes of these motor domains are not the major determinants of the directionality of their movement along microtubules.
Fujiwara
S
S
Cardiovascular Research Institute, University of California, San Francisco 94143, USA.
Kull
F J
FJ
Sablin
E P
EP
Stone
D B
DB
Mendelson
R A
RA
D000818:Animals; D003001:Cloning, Molecular; D004330:Drosophila / Q000201:enzymology; D029721:Drosophila Proteins; D004926:Escherichia coli; D006801:Humans; D016547:Kinesins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008868:Microtubule Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008970:Molecular Weight; D009502:Neutrons; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012542:Scattering, Radiation
D029721:Drosophila Proteins; D008868:Microtubule Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C066696:ncd protein, Drosophila; D016547:Kinesins