Fig. 4.

Stopped-flow measurements of light scattering to examine the early stages of clathrin cage disassembly. Clathrin cages (0.09 μM triskelia) premixed with 0.1 μM auxilin were mixed with Hsc70 (concentrations in μM shown on graph) and 500 μM ATP, and perpendicular light scattering was measured using stopped-flow techniques. The closely spaced raw data was reduced to a frequency of 3.3 s^-1 (closed circles) and fitted (dashed lines) to a system of first-order ordinary differential equations corresponding to the reaction mechanism shown in Scheme 1, using the software DYNAFIT (29, 30). Data corresponding to 0 and 0.25 μM Hsc70 were omitted from the fit because of their low information content, as was data collected after 12 s.