Influence of polymer molar mass and mixture stoichiometry on polyelectrolyte complexes of poly(L-arginine) and Poly(L-glutamic acid)

Poly(L-arginine) (PARG) and poly(L-glutamic acid) (PLGA) homopolypeptides were custom synthesized by precision N-carboxyanhydride ring-opening polymeri-zation methods with two molar masses, matched for pairs of cationic and anionic polypeptides (degrees of polymerization n = 100 and n = 500). The conformations of the homopolypeptides were probed using circular dichroism (CD) and FTIR spectroscopy which revealed the presence of mainly polyproline II (PPII) conformation. Small-angle X-ray scattering (SAXS) showed concentration-dependent polyelectrolyte peaks and form factor with high q scaling due to the excluded volume behaviour of the wormlike chains. We then examined polyelectrolyte complexation in mixtures of pairs of PARG and PLGA polypeptides with matched molar masses. Precipitation was generally observed and the structures of precipitates, supernatant and resuspended precipitates were investigated using CD, SAXS and cryo-TEM. These revealed that, contrary to prior suggestions in the literature, the precipitates contain mostly polypeptides in a PPII-like conformation, and there is only a minimal beta-sheet content (which is enhanced upon drying the sample during preparation for certain measurements). The precipitates have a fractal-like structure as revealed by cryo-TEM and SAXS. Our findings on the structure of polypeptide complex precipitates contribute to the understanding of phase separation of poly -electrolyte complexes and coacervation and may shed light on the formation of inter-cellular bodies of proteins and peptides such as Lewy and other inclusion bodies.

Automated summary

This study investigated the unique structures of PARG and PLGA polypeptides using various analytical methods, revealing that they mainly adopt PPII conformation and form fractal-like precipitate structures. Understanding the structure of polypeptide complex precipitates contributes to the comprehension of phase separation in polyelectrolyte complexes and coacervation, and sheds light on the formation of inter-cellular bodies of proteins and peptides.