Molecular Basis of Filamin A-FilGAP Interaction and Its Impairment in Congenital Disorders Associated with Filamin A Mutations
Figure 7
Effect of disease-related mutations in repeat 23 of FLNa on FilGAP binding.
(A) Amino acid sequence of human IgFLNa23. Point (L2439M) and deletion (Ä7 and Ä3) mutants of FLNa are linked to human diseases and shown in underline. Box arrows indicate â-sheet strands. (B) Rheological properties of the FLNa mutants. Left panel, Apparent viscosity of 24 ìM actin polymerized with the indicated molar ratios of wild-type and mutant FLNa. Error bars indicate SEM from triplicates. Right panel, Maximum shear stress supported by 12 ìM F-actin polymerized with or without 0.06 ìM wild-type and mutant FLNa. Error bars indicate s.d. from triplicates. The linear and nonlinear rheological properties are provided in (Figure S6). (C) In vitro binding of recombinant human FLAG-FLNa and FilGAP purified from Sf9 insect cells. FilGAP was incubated with increasing amounts of wild-type or mutant FLNa and the complex was co-immunoprecipitated with FLAG-specific mAb immobilized on agarose beads. Bound proteins were detected by immunoblotting.