The kinetics of aggregation of poly-glutamic acid based polypeptides

The aggregation of two negatively-charged polypepticles, poly-L-glutamic acid (PE) and a copolymer of polyglutamic acid and poly-alanine (PEA), has been studied at different pepticle and salt concentrations and solution pH conditions. The kinetics of aggregation were based on Thioflavin T (ThT) fluorescence measurements. The observed lag phase shortened and the aggregation was faster as the pH approached the polypepticles'isoelectric points. While the initial polypepticle structures of PE and PEA appeared identical as determined from circular dichroism spectroscopy, the final aggregate morphology differed; PE assumed large twisted lamellar structures and the PEA formed typical amyloid-like fibrils, although both contained extensive P-sheet structure. Differences in aggregation behavior were observed for the two polypeptides as a function of salt concentration; aggregation progressed more slowly for PE and more quickly for PEA with increasing salt concentration. Several models of aggregation kinetics were fit to the data. No model yielded consistent rate constants or a critical nucleus size. A modified nucleated polymerization model was developed based on that of Powers and Powers [E.T. Powers, D.L. Powers, The kinetics of nucleated polymerizations at high concentrations: Amyloid fibril formation near and above the "supercritical concentration", Biophys.J. 91 (2006) 122-132], which incorporated the ability of oligomeric species to interact. This provided a best fit to the experimental data. (c) 2008 Elsevier B.V. All rights reserved.