Temperature dependence of the structure of aggregates of tobacco mosaic virus protein at pH 7.2. Static synchrotron small-angle X-ray scattering.
The small-angle X-ray scattering (SAXS) method using a synchrotron radiation source was applied to the study of the self-aggregation process of tobacco mosaic virus protein (TMVP) at a concentration of 5.0 or 12.0 mg ml-1 in 50 mM or 100 mM-phosphate buffer (ionic strengths approx. 0.1 and 0.2, respectively) at pH 7.2 in the temperature region of 4.8 to 25.0 degrees C. This paper presents the results of static measurements of SAXS. Sedimentation velocity experiments were performed simultaneously under the same conditions. These results are qualitatively parallel to those of the SAXS measurements, although the size of stacked disks derived from the SAXS measurements is larger than that derived from the sedimentation experiments, suggesting a change in the equilibrium conditions in the centrifugal field. Qualitative analysis of the SAXS data with model simulation calculations implies that the aggregation of TMVP consists of two steps: (1) the aggregation of A-protein comprising a few subunits to form double-layered disks; and (2) the random polymerization of double-layered disks by disk-stacking. Increase in temperature, ionic strength or protein concentration induced TMVP to polymerize to form a double-layered disk or a quadruple-layered short rod with consumption of A-proteins, accompanied by a small number of multi-layered short rods. The SAXS results indicate that the A-protein and the multilayered short rods are polydisperse with respect to size and shape, i.e. the mixture of A-protein, double-layered disks and multi-layered short rods coexists in the equilibrium state without pressure-induced partial dissociation of TMPV as observed during normal ultracentrifugation, and even under solution conditions in which the formation of double-layered disks or higher-order aggregates is favored.
Hiragi
Y
Y
Institute for Chemical Research, Kyoto University, Japan.
Inoue
H
H
Sano
Y
Y
Kajiwara
K
K
Ueki
T
T
Kataoka
M
M
Tagawa
H
H
Izumi
Y
Y
Muroga
Y
Y
Amemiya
Y
Y
D036022:Capsid Proteins; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D008954:Models, Biological; D008970:Molecular Weight; D013696:Temperature; D014027:Tobacco Mosaic Virus; D014764:Viral Proteins; D014961:X-Ray Diffraction
D036022:Capsid Proteins; C009496:H protein, Tobacco mosaic virus; D046911:Macromolecular Substances; D014764:Viral Proteins