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

• DOI: 10.1016/j.polymer.2022.125497
• Publication Date: 2022-11-14T17:30:19Z
• Authors: Valeria Castelletto; Lucas de Mello; Foteini Arfara; Hermis Iatrou; Jani Seitsonen; Ian W. Hamley
• Keywords: Polyproline helix; Small-angle X-ray scattering; Molar mass; Lyotropic; Coacervate; Dispersity

Funding Information: This work was supported by EPSRC Fellowship grant EP/V053396/1 to IWH , and by the European Union and Greek national funds through the Operational Program "Competitiveness, Entrepreneurship and Innovation" , under the call “ RESEARCH-CREATE-INNOVATE ” (project code: Τ1ΕΔΚ-01833 ). We are grateful for the award of synchrotron SAXS beamtime at SOLEIL (proposal no. 20211309 ) and the support of Thomas Bizien (SOLEIL) and Elisabetta Rosa ( University of Naples Federico II, Italy ) during the measurements. Beamtime at Diamond was awarded under reference SM29895-1 and we thank Nikul Khunti for assistance. We acknowledge access to instruments of the Chemical Analysis Facility at the University of Reading. Funding Information: This work was supported by EPSRC Fellowship grant EP/V053396/1 to IWH, and by the European Union and Greek national funds through the Operational Program "Competitiveness, Entrepreneurship and Innovation", under the call “RESEARCH-CREATE-INNOVATE” (project code: Τ1ΕΔΚ-01833). We are grateful for the award of synchrotron SAXS beamtime at SOLEIL (proposal no. 20211309) and the support of Thomas Bizien (SOLEIL) and Elisabetta Rosa (University of Naples Federico II, Italy) during the measurements. Beamtime at Diamond was awarded under reference SM29895-1 and we thank Nikul Khunti for assistance. We acknowledge access to instruments of the Chemical Analysis Facility at the University of Reading. Publisher Copyright: © 2022 The Authors<br/>Peer reviewed<br/>Poly(L-arginine) (PARG) and poly(L-glutamic acid) (PLGA) homopolypeptides were custom synthesized by precision N-carboxyanhydride ring-opening polymerization 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 β-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 polyelectrolyte 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.<br/>