Charge state of arginine as an additive on heat-induced protein aggregation

July 28, 2017


Charge state of arginine as an additive on heat-induced protein aggregation


Takumi Miyatake, Shunsuke Yoshizawa, Tsutomu Arakawa, Kentaro Shiraki




International Journal of Biological Macromolecules


Arginine (Arg) is one of the most versatile solvent additives, such as suppressing protein aggregation, increasing solubility of small aromatic compounds and peptides, and preventing protein binding on solid surfaces. In this study, we investigated the role of the charged state of α-amino group of Arg for the prevention of protein aggregation. As expected, Arg effectively suppressed thermal aggregation of hen egg-white lysozyme at neutral pH, whereas the suppression effect diminished at and above pH 9.0, which corresponds to the pK of Arg’s α-amino group. The pH dependence of Arg as an aggregation suppressor was confirmed by additional experiments with neutral proteins, bovine hemoglobin and bovine γ-globulin. Interestingly, N-acetylated arginine, which lacks the α-amino group, showed a weaker suppressive effect on protein aggregation than Arg, even at neutral pH. These results indicate that both positively charged α-amino group and guanidinium group play important roles in suppressing heat-induced protein aggregation by Arg. The elucidated limitation of Arg at alkaline pH provides new insight in the application as well as the mechanism of Arg as a solvent additive.




Circular dichroism, Tertiary structure, Protein folding, Thermodynamics, Aggregation, Thermal stability, Protein denaturation, Biochemistry