TtClpP (3 μM) in a buffer containing 100 mM tris (pH 8.5), 100 mM NaCl, and 5% glycerol without or with 5 mM added bortezomib was measured using a DynaPro NANOSTAR (Wyatt, Santa Barbara, USA) instrument operated at 25° or 45°C and analyzed with the associated DYNAMICS software. Each experiment was performed in triplicate (three times for each of two TtClpP samples, both measured at 25° and 45°C), and hydrodynamic radii were calculated as the average of these replicates. Theoretical hydrodynamic radii of compressed TtClpP [based on PDB: 3QWD (18)] and extended TtClpP (using the TtClpP:bortezomib structure), after modeling of missing residues in the N-terminal β-hairpins and the N and C termini using SWISS-MODEL (63), were calculated using Hydropro10 (64) using a solvent viscosity and a solvent density of 0.0010387 poise and 1.0114 g/cm3, respectively (5% glycerol, 25°C), or 0.00068656 poise and 1.0043 g/cm3, respectively (5% glycerol, 45°C).

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