Figure 2
Interaction of the ectodomains of LRP10 and APP. (A) Schematic representation of the LRP10 FLAG-tagged deletion mutants used to determine the binding domain of APP. The structural elements of LRP10 are depicted, including (from amino to carboxyl terminus) the CUB domains, LDLRA repeat, and transmembrane (TM) domain. CD, Cytoplasmic domain; ED, Ectodomain. (B) The interaction between LRP10 and APP does not depend on the cytoplasmic domain of LRP10. Lysates of HEK cells transfected with APP695 and pcDNA3-FLAG, FLAG-tagged LRP10ΔCD, or FLAG-tagged LRP10ΔED were immunoprecipitated with anti-APP or anti-FLAG and were immunoblotted with anti-APP or anti-FLAG antibody. (C) Schematic representation of the GST-APP deletion mutants used to determine the binding domain of LRP10. The structural elements of APP are depicted, including the cysteine, acidic, carbohydrate, amyloid beta, and cytoplasmic domains. (D) In vitro interaction of LRP10 with the ectodomain of APP. The APP deletion mutants shown in (C) and the GST protein (10 μg each) were immobilized on glutathione beads and were incubated with in vitro translated 35 S-labeled LRP10. Bound proteins were separated by SDS-PAGE and were detected by autoradiography. GST proteins were detected by coomassie staining. Input equaled 2.5% of the total in vitro-translated product.