DeltaV Change Research Articles

The thermodynamic analysis of protein stabilization by sucrose and glycerol against pressure-induced unfolding.

We have studied the reaction native left arrow over right arrow denatured for the 33-kDa protein isolated from photosystem II. Sucrose and glycerol have profound effects on pressure-induced unfolding. The additives shift the equilibrium to the left; they also cause a significant decrease in the standard volume change (DeltaV). The change in DeltaV was related to the sucrose and glycerol concentrations. The decrease in DeltaV varied with the additive: sucrose caused the largest effect, glycerol the smallest. The theoretical shift of the half-unfolding pressure (P1/2) calculated from the net increase in free energy by addition of sucrose and glycerol was lower than that obtained from experimental mea- surements. This indicates that the free energy change caused by preferential hydration of the protein is not the unique factor involved in the protein stabilization. The reduction in DeltaV showed a large contribution to the theoretical P1/2 shift, suggesting that the DeltaV change, caused by the sucrose or glycerol was associated with the protein stabilization. The origin of the DeltaV change is discussed. The rate of pressure-induced unfolding in the presence of sucrose or glycerol was slower than the refolding rate although both were significantly slower than that observed without any stabilizers.

Photosynthetic Electron Carriers at a Heptane-Water Interface

Surface properties of monomolecular films of chlorophyll (Chl), plastocyanin (Pc), cytochrome c (Cyt) and ferridoxin (Fd) were measured at a heptane-water interface. Mixed films of Chl and the other components were examined in darkness and in light. The area/molecule, A, for Fd (from Cl. pasteurianum) is much larger at a heptane-water than Fd (from spinach) at an air-water interface. This difference in A may be the source of the Fd or the extent of denaturation at the different interfaces. There appears to be a photoreaction between Chl and Fd in the presence of ascorbate. The A for Cyt is much larger at a heptane-water than at an air-water interface. In mixed films there is a strong interaction between Chl and reduced Cyt in the dark. No such interaction is observed between Chl and oxidized Cyt. With mixed films of Chl and reduced Pc there is a significant, reversible, light induced change in deltaV.

Interactions between retinal and phospholipids in monomolecular films at acid pH.

The surface properties of mixed monomolecular films of retinal and phospholipids (p. lipids) are measured as a function of mole fraction at a nitrogen-water interface. An acid pH of 6.0 is maintained in the aqueous phase. Before irradiation the surface potential deltaV for 9-cis retinal, 11-cis retinal, phosphatidyl serine (PS) and phosphatidyl ethanolamine (PE), at pi=12 dyn/cm, are 490 mV, 645 mV, respectively. Before irradiation, A0 for 9-cis and 11-cis are 58 A2 and 48 A2, respectively. Experimentally measured isotherms are compared with theoretically calculated isotherms. In case of mixed films of retinal and PS the experimental isotherms are greater than theoretical, while mixed films of retinal and PE are smaller than theoretical. A maximum value for the difference between theoretical and experimental areas are obtained at (retinal)/(p. lipid)=0.1. Retinal and p. lipid do not appear to form a Schiff base, charge transfer or any other type of complex at pH 6. A eutectic type mixture between retinal and p. lipid may occur on the surface. A light induced change in deltaV of -130 mV is observed in the case of 11-cis and PE. The significance of these findings with respect to visual excitation is considered.