Charge-induced low-temperature gelation of mixed proteins and the effect of pH on the gelation: A spectroscopic, rheological and coarse-grained molecular dynamics study

We report the gelation of mixed proteins consisting of oppositely charged lysozyme and serum albumins at various pH. The results from rheological tests showed that at a pH of 7, the gelation temperature (T_gel) of the oppositely charged proteins was lower than the melting temperature (T_m) of the individual protein. To ascertain the conformational state of the proteins at the observed T_gel, the attenuated total-reflectance Fourier-transform infrared (ATR-FTIR) spectra of the proteins were acquired. The recorded spectra showed that the proteins were predominantly alpha helical, suggesting that the observed gelation was electrostatically triggered. However, as the solution pH was changed to acid or alkaline regime, all the proteins became similarly charged and showed T_gel < T_m which was attributed to pH-induced denaturation. Surprisingly, however, the serum albumins were remarkably stable at the alkaline pH of 9 and 10 but very labile at the acidic pH. In contrast, the LYZ was more stable at the acidic than alkaline pH. To understand the role of the opposite charges in the gelation, coarse-grained molecular dynamics (CGMD) simulations revealed an increase in the aggregation of the oppositely charged proteins compared with the pure or similarly charged protein mixture.