Effects of Group 3 LEA protein model peptides on desiccation-induced protein aggregation

Group 3 late embryogenesis abundant (G3LEA) proteins have amino acid sequences with characteristic 11-mer motifs and are known to reduce aggregation of proteins during dehydration. Previously, we clarified the structural and thermodynamic properties of the 11-mer repeating units in G3LEA proteins using synthetic peptides composed of two or four tandem repeats originating from an insect (Polypedilum vanderplanki), nematodes and plants. The purpose of the present study is to test the utility of such 22-mer peptides as protective reagents for aggregation-prone proteins. For lysozyme, desiccation-induced aggregation was abrogated by low molar ratios of a 22-mer peptide, PvLEA-22, derived from a P. vanderplanki G3LEA protein sequence. However, an unexpected behavior was noted for the milk protein, alpha-casein. On drying, the resultant aggregation was significantly suppressed in the presence of PvLEA-22 with its molar ratios > 25 relative to alpha-casein. However, when the molar ratio was < 10, aggregation occurred on addition of PvLEA-22 to aqueous solutions of alpha-casein. Other peptides derived from nematode, plant and randomized G3LEA protein sequences gave similar results. Such an anomalous solubility change in alpha-casein was shown to be due to a pH shift to ca. 4, a value nearly equal to the isoelectric point (pI) of alpha-casein, when any of the 22-mer peptides was mixed. These results demonstrate that synthetic peptides derived from G3LEA protein sequences can reduce protein aggregation caused both by desiccation and, at high molar ratios, also by pH effects, and therefore have potential as stabilization reagents. (C) 2012 Elsevier B.V. All rights reserved.