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Arborescent Poly(l-glutamic acid)s as Standards To Study the Dense Interior of Polypeptide Mesoglobules by Pyrene Excimer Fluorescence

A series of arborescent poly(l-glutamic acid)s of generations 0 to 3 (PGA(GY) with Y = 0–3) were randomly labeled with 1-pyrenemethylamine to yield several Py-PGA(GY) constructs with pyrene contents ranging between 2.5 and 22 mol %. The density (ρ) of the interior of the PGA(GY) samples was estimated in N,N-dimethylformamide (DMF) and dimethyl sulfoxide (DMSO) by conducting gel permeation chromatography and dynamic light scattering experiments to determine their molar mass and hydrodynamic diameter, respectively. It was determined that ρ increased with the generation number from PGA(G1) to PGA(G2), which promoted more contacts between the pyrene labels. The increase in the number of pyrene–pyrene contacts was quantified with the parameter Nblob obtained by analysis of the fluorescence decays for the Py-PGA(GY) samples in DMF and DMSO. In the analysis, Nblob represented the number of structural units, i.e., glutamic acid residues, comprised inside the volume probed by an exited pyrene, termed a blob. Insid...