High pressure effects on the endothermic association of tobacco mosaic virus protein

Abstract

Tobacco mosaic virus protein in phosphate buffer pH 6.5–7.0 (I=0.1 M) shows endothermic polymerization accompanied by water release of the capsomers. At protein concentrations c ∼ 2 mg/ml the transition temperature is T ^*=20 ± 1‡ C. As indicated by the increase of the partial specific folume (δV ₂=0.0049 ± 0.0003 cm³/g) in going from A-protein to helical rods at pH 6.50, the assembly reaction is expected to be inhibited by high pressure; the corresponding isobars of the endothermic polymerization should be shifted to higher T ^* values.

Turbidity measurements at pressures 1<p<1,500 bar are in agreement with the given hypothesis: both, double discs and helical rods are found to be dissociated at elevated pressure, the latter showing somewhat higher stability. At 700 bar the transition temperature of helix formation is shifted by 14‡ C to higher temperatures.

Complete reversibility of the pressure dependent dissociation-association without “hysteresis” proves the process to represent a true equilibrium. At low temperatures and high pressures the association equilibrium is shifted to a molecular weight distribution with M _w< M (A-protein). Increased co operativity in the transition A-protein → helical rods, as well as an apparent inversion of the sign of the reaction volume at high temperatures and pressures are caused by pressure induced pH shifts. Adjusting the pH at high pressure to the value at ambient pressure allows to eliminate both effects.

The product of association at high pressure differs in its conformation from the end product obtained from the endothermic polymerization at 1 bar and subsequent pressure application.