A specific effect of calcium ion on the polymerization-depolymerization of tobacco mosaic virus protein.

Tobacco mosaic virus protein stored in the cold at low ionic strength between pH 5 and 5.5 is highly polymerized. When such protein is brought to room temperature and mixed with acetate buffer and additional electrolyte to give a final pH of 6.5 and ionic strength of 0.1, the protein is still in the polymerized state. When the temperature is dropped to about 5 °C, the protein depolymerizes rapidly, in the normal manner, if the added electrolyte is barium chloride, magnesium chloride, or potassium chloride. However, if it is 0.01 m calcium chloride, the depolymerization is slow, requiring about 12 h to reach completion. When the temperature of this depolymerized solution is raised, the protein polymerizes rapidly; when the temperature is dropped, the protein depolymerizes rapidly, just as in solutions free of calcium.Ion-binding studies show that calcium is bound to the protein during the initial step when it is brought to pH 6.5 and room temperature in the presence of calcium. The calcium is released during the slow depolymerization when the temperature is dropped and is not bound again during polymerization at pH 6.5, brought about by an increase in temperature. This means that polymerized protein at pH 5.5 has a structure capable of binding calcium ions, probably a helical structure like that of the protein in the virus. When pH is raised to pH 6.5 at room temperature, this structure remains long enough for calcium to be bound when present. These calcium ions stabilize the polymer, resulting in slow depolymerization when the temperature is lowered. When the temperature is raised at pH 6.5, a different, looser polymer structure is obtained, one not capable of binding calcium.