Figure 8.

Determination of the Binding Affinity of cpSRP54/uL4 Complex Formation Using Microscale Thermophoresis.

(A) to (F) Fluorescently labeled His-cpSRP54M (see [A], and [D] to [F]), His-cpSRP54MΔ415-418 (B), and His-cpSRP54MΔ536-540 (C) (20 nM) were titrated with His-uL4 (see [A] to [C]) or the indicated His-uL4 constructs (see [D] to [F]). The difference in normalized fluorescence was plotted against the concentration of the indicated uL4 constructs. Raw microscale thermophoresis traces are shown in Supplemental Figure 7. The K_d values and corresponding sds were calculated from three independent replicates. The recombinant proteins used for microscale thermophoresis are shown in Supplemental Figure 8.