Fig. 2.

The intermediate mimic is a monomer. (a), Gel filtration (Sephadex G-75) of the intermediate mimic (filled squares, repeated three times at protein concentrations from ~10 μM to ~100 μM) and standards (circles), including horse myoglobin (17 kDa), chicken ovalbumin (44 kDa), bovine γ-globulin (158 kDa), and bovine thyroglobulin (670 kDa). The arrows indicate the anticipated monomeric and dimeric positions. (b), Longitudinal and transverse relaxation times, T₁ (square) and T₂ (circle). The straight lines illustrate the values of 0.55 s and 75 ms respectively. (c), c(s) distributions based on sedimentation velocity data collected at 60 krpm, 300 nm and 20.0°C are shown for loading concentrations of 100 μM (red) and 200 μM (green). The inset shows the c(s) distributions for sedimentation velocity data collected at 60 krpm, 285 nm, 20.0°C and loading concentrations of 7.1 μM (red), 14.3 μM (green) and 28.6 μM (blue). (d), Overlay of the ¹H-¹⁵N HSQC spectra of the intermediate mimic at 1 mM (black) and 200 μM (red). The peaks in black were slightly shifted toward right to help to observe them.