Fig. 4.

Vinculin Vd1 binding analysed by gel filtration. Vinculin Vd1 was incubated with (A) the talin 4-helix or (B) the 5-helix polypeptide at various temperatures, and complex formation was analysed on a gel filtration column at room temperature (RT). (A) Incubation of the 4-helix with Vd1 resulted in complex formation at room temperature (dotted line). (B) The 5-helix bundle did not bind vinculin Vd1 at room temperature (solid line), but pre-incubation of the proteins at 37 °C increased complex formation (broken line) and pre-incubation at 45 °C resulted in predominantly complexed proteins (dotted line).