Light Diffraction of in vitro Crystals of Six Tobacco Mosaic Viruses

Abstract

Iridescent gels of the common, tomato mosaic, para-tobacco mosaic, ribgrass mosaic, sunnhemp mosaic, and cucumber 4 tobacco mosaic virus strains were prepared using the purification method of Boedtker and Simmons (J. Amer. Chem. Soc. 1958). Macrocrystals which were stable for many months could be grown from iridescent gels of all viruses when stored at 0-10 °C. The gels exhibited various structural phases (nematic and smectic) differing in density. The structure of the microcrystalline gels and of macrocrystals was investigated by means of optical diffraction. Distinct Bragg reflections were observed from which a mean periodicity of 340 nm was derived for all virus strains. The results indicate a crystalline multilayer structure as first proposed by Oster (J. Gen. Physiol. 33, 445, [1950]). The angular positions and widths of the Bragg reflections depended on the concentration of virus and the ionic strength. The periodicity decreased from 365 nm at 30 mg/ml virus to about 315 nm at 109 mg/ml virus, i.e. the periodicity approached the 300 nm length of the virion. Model calculations taking into account Debye-Hückel repulsive and van der Waals attractive forces gave a qualitative explanation for the crystalline structure and its observed concentration dependence. The in vitro crystals resemble immature crystal forms observed in the living cell.