RasD binding does not depend on any single repeat in the rod domain nor the actin binding domain. Western blot using the antiRasD antibody to detect RasD in immunoprecipitations using protein extract from HPF616 (lane 1, WT) (AX2 ectopically expressing RasD), HPF631 (lane 2, Null) (HG1264 ectopically expressing RasD) and strains HPF632-638 (lanes 3–10) (HG1264 ectopically expressing RasD and one of filaminΔ1–6 or filaminΔABD as indicated). RasD is detected in all IP samples with the exception of the null mutant HPF631, indicating that filamin is required to pull down RasD and RasD interacts with filamin regardless of which portion of the scaffolding protein is absent.
Annesley et al. BMC Cell Biology 2007 8:48 doi:10.1186/1471-2121-8-48