Fig 1: FMNL1 Is a High-Affinity Cargo for UNC119A(A) Western immunoblot where 30-μg GST-tagged full-length UNC119A was used to pull down Jurkat lysate in the presence or absence of 30-μg ARL2GppNHp or ARL3GppNHp.(B) Full-length UNC119A was titrated into a solution of 0.2-μM fluorescein-labeled myristoylated FMNL1 (○) or FMNL3 (⋅) peptides. Binding affinities for FMNL1 and FMNL3 were calculated as 10.1 ± 3.5 nM and 856 ± 156 nM, respectively.(C and D) Fluorescence polarization measurement where 0.5-μM full-length UNC119A was added to 0.5-μM fluorescein-labeled myristoylated FMNL1 (C, Myr-GNAAGSAEQPAG) or FMNL3 (D, Myr-GNLESAEGVPGE) peptide followed by 5-μM GppNHp-loaded ARL2 or ARL3.(E) GppNHp-loaded ARL2 and ARL3 were added in 2.5-μM steps to a mixture of 0.5-μM fluorescein-labeled myristoylated FMNL1 peptide and 0.5-μM UNC119A as indicated.