Figure 3. CG-MALS for SKp and CaM. (A–D)... | Rockefeller University Press

Figure 3.

$Figure 3. CG-MALS for SKp and CaM. (A–D) Black symbols show the time-averaged light scattering signal from multiple detectors as plotted versus protein composition. The composition varies with the increasing fraction of CaM (orange horizontal scale) and the decreasing fraction of SKp (black horizontal scale). Data in A and B are experiments in zero Ca2+ from the self+hetero-association protocol (triangles). Data in C and D are at saturating Ca2+ from both self+hetero-association (triangles) and dual crossover experiments (circles). All experiments in A and B were fitted simultaneously. The fits are represented by the gray broken lines. The maximum concentrations of CaM and SKp are 1.3 µM and 1.6 µM, respectively, in A for “low protein concentration,” and 2.7 µM and 4.2 µM, respectively, in B for “high protein concentration.” Data in C and D were also fit simultaneously. The maximum concentrations are 0.8 µM CaM and 1.6 µM SKp in C and 2.6 µM CaM and 3.8 µM SKp in D. The black closed circles distributed around the solid gray lines are the residuals to the fits. (E–H) The modeled contributions of each molecular species to the light scattering signal in A–D, respectively; the fitted curves are reproduced as broken gray lines and the vertical and horizontal axes are the same as in A–D. In E and F, the modeled contribution of each molecule to the light scattering curves are: SKp, solid black; CaM, solid orange; 2SKp/1CaM, green; 1SKp/1CaM, blue; incompetent SKp, broken black; incompetent CaM, broken orange. In G and H, the components are the same, but required the addition of a 1SKp/2CaM (red) component.$

CG-MALS for SKp and CaM. (A–D) Black symbols show the time-averaged light scattering signal from multiple detectors as plotted versus protein composition. The composition varies with the increasing fraction of CaM (orange horizontal scale) and the decreasing fraction of SKp (black horizontal scale). Data in A and B are experiments in zero Ca²⁺ from the self+hetero-association protocol (triangles). Data in C and D are at saturating Ca²⁺ from both self+hetero-association (triangles) and dual crossover experiments (circles). All experiments in A and B were fitted simultaneously. The fits are represented by the gray broken lines. The maximum concentrations of CaM and SKp are 1.3 µM and 1.6 µM, respectively, in A for “low protein concentration,” and 2.7 µM and 4.2 µM, respectively, in B for “high protein concentration.” Data in C and D were also fit simultaneously. The maximum concentrations are 0.8 µM CaM and 1.6 µM SKp in C and 2.6 µM CaM and 3.8 µM SKp in D. The black closed circles distributed around the solid gray lines are the residuals to the fits. (E–H) The modeled contributions of each molecular species to the light scattering signal in A–D, respectively; the fitted curves are reproduced as broken gray lines and the vertical and horizontal axes are the same as in A–D. In E and F, the modeled contribution of each molecule to the light scattering curves are: SKp, solid black; CaM, solid orange; 2SKp/1CaM, green; 1SKp/1CaM, blue; incompetent SKp, broken black; incompetent CaM, broken orange. In G and H, the components are the same, but required the addition of a 1SKp/2CaM (red) component.