FIG. 5.

(A) EPR spectra of the Eg5 dimer under 32 different experimental conditions. (B) Experimentally determined basis functions from [16]. (C) Comparison of immobile fraction determined using a linear least-squares method [17] with the basis functions in (B) to those obtained from constrained two-state decomposition (D). The experimentally derived basis spectra were also decomposed and are shown as triangles. The straight line shows equal population estimates from both methods. Procedures for expression and purification of Eg5 protein, conjugation of the MSL probe, sample preparation, and acquisition of EPR data are described in [16].