Assessment of Rab11-FIP2 interacting proteins in vitro.

Members of the Rab family of small GTPases are involved in multiple trafficking events in both endocytotic and biosynthetic pathways. To understand more fully the regulation of these events, a concerted effort is underway to ascertain the binding partners and regulators of Rabs. Here, we describe methods to assess binding of Rab11a with Rab11-FIP2 and other Rab11-FIPs utilizing a modified far-Western approach. We then broaden this application to assess binding of Rab11-FIP2 with myosin Vb and homodimerization of Rab11-FIP2.