Abstract
Clathrin labeled with eosin maleimide showed physicochemical properties similar to the native clathrin. The diffusion coefficients of clathrin protomers and cages measured at 20°C by the fluorescence recovery after photobleaching technique (FRAP) were found equal to (9 ± 1) · 10 −8cm 2·s −1 and (3 ± 0.4) · 10 −8cm 2 ·s −1, respectively. After incubation with isolated mouse liver nuclei suspended in an aqueous buffer, FRAP measurements showed that 78% of clathrin was immobilized on the nuclear surface. This immobile fraction might correspond to aggregates of molecules resembling coated pits. The mobile fraction had a diffusion coefficient of 2.5 · 10 −9cm 2·s −1 which was reduced seven times when the suspension medium of the nuclei contained 50% sucrose, showing that the aqueous phase exerted an important drag on the clathrin molecules motion diffusing on the nuclear surface.
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