An Effective Method for Determining the Degree of Oli-gomerization of hnRNPA2 Low Complexity Domain

The following files include data presented in the manuscript "An Effective Method for Determining the Degree of Oligomerization of hnRNPA2 Low Complexity Domain written by Paulina Żeliszewska, Zbigniew Adamczyk, Pooja Shah, Anna Kluza, Aneta Michna, and Anna Bratek-Skicki.

Figure 1. The nominal charge (Q) of the chicken-lysozyme molecule (◊) and the hnRNPA2 LCD monomer (●) versus pH calculated using the PROPKA 3.0 algorithm. 

Figure 3. Dependence of the zeta potential (●) (left-hand axis) and the electrophoretic mobility (◊) of the hnRNA2 LCD molecules on pH determined by the LDV method in 10 mM NaCl. The line is the guide for the eyes.

Figure 4. AFM micrographs of hnRNPA LCD protein layers on mica after 15 minutes of adsorption. Experimental conditions: bulk protein concentration c_b = 0.5 mg L^-1, pH 3.5, 10 mM NaCl. (a) Freshly prepared protein solution; (b) Solution prepared from protein stored for over 12 months.

Figure 5. Histogram of hnRNPA2 LCD protein oligomers size distribution on mica (adsorption conditions as in Figure 4). (a) Freshly prepared protein solution, average oligomer size 11 ± 4 nm; (b) Solution prepared from protein stored for over 12 months at -70 C, average oligomer size 21 ± 5 nm.

Figure 6. The dependence of the zeta potential of polymer microparticles (PS800) on the hnRNPa2 LCD concentration; adsorption conditions: protein volume 5 ml, particle suspension volume 5 ml, particle concentration 100 mg L^-1, pH 4.0, 10 mM NaCl. (a) Freshly prepared protein solution (stock suspension concentration of 50 mg L^-1 ); the triangle points show the experimental results obtained from the LDV measurements (stock suspension concentration of 50 mg L^-1 ), the circles show the experimental points (stock suspension concentration of 125 mg L^-1 ), the solid lines show the theoretical results calculated for adsorption of the monomer (red line 1) and oligomer composed of 13 molecules (green line 2 );(b) Stored protein, the points denote experimental results obtained from the LDV measurements, the solid lines show theoretical results calculated for adsorption of the monomer (red line 1), oligomer composed of 13 molecules (green line 2 ), and the oligomer composed of 64 molecules (blue line 3). 

Figure 7. The dependence of the zeta potential of polymer microparticles (PS800) on the bulk hnRNPA2 LCD concentration, pH 4.0, 150 mM NaCl, (a) Freshly prepared protein solutions (stock concentration 125 mg L^-1) the points denote experimental results obtained from the LDV measurements, the solid line shows the theoretical results calculated for adsorption of the oligomer composed of 64 molecules; (b) Solutions prepared from stored stock suspensions, the points denote experimental results obtained from the LDV measurements, the solid lines show the theoretical results calculated for adsorption of the oligomer composed of 110 molecules. 

Figure 8. Stability of hnRNPA2 LCD functionalized particles determined by the DLS measurements. (a) pH 4.0, 10 mM NaCl, and by the electrophoretic mobility measurements; (b) pH 4.0, 10 mM NaCl; (c) pH 7.4, 10 mM NaCl.

Figure 9. Dependence of the zeta potential of the PShnRNPA2 LCD particles on pH. The points represent experimental data acquired using the LDV method for 10 mM NaCl of freshly prepared solutions (stock suspension concentration 125 mg L ^-1). The solid line is the guide for the eyes.