Fig. 3.

Identification of connector binding region of scaffolding protein. A, the ability of scaffolding deletion mutants to solubilize connector protein at low ionic strength buffer was assayed by SDS-PAGE following centrifugation. The amounts of input and insoluble protein are shown in the odd and even lanes, respectively. Lanes 1 and 2, connector protein only; lanes 3 and 4, connector plus wild type (WT) scaffolding protein; lanes 5 and 6, connector plus Δ74–98 scaffolding protein; lanes 7 and 8, connector plus Δ70–98 scaffolding protein. B, the effect of temperature on the ability of wild type and temperature-sensitive mutant scaffolding protein to solubilize connector protein. The fraction of connector solubilized following incubation at 20, 30, and 42 °C was quantified by centrifugation and SDS-PAGE for connector protein alone (closed squares) or connector in the presence of wild type (open squares) or S65N scaffolding protein (open triangles).