EhP3, a homolog of 14-3-3 family of protein participates in actin reorganization and phagocytosis in Entamoeba histolytica
Fig 6
(A) Graph depicts analysis of G-actin solid phase plate binding assay for EhP3. The histogram represents relative mean intensity of fluorescence observed on binding to actin. CaBP1 was used as binding control. (B) Immunoblot analysis of the F-actin co-sedimentation assay for EhP3. Purified EhP3 protein incubated at different concentration (2.5, 5, 10 μM) with polymerised G-actin (5 μM) or without F-actin were separated into supernatant (S) and pellet (P) fractions by high speed ultracentrifugation and analysed by SDS PAGE. (S-supernatant, P-pellet). (C) Model summarising reported/predicted interactions between 14-3-3 and actin regulatory proteins that controls cell cytoskeleton remodelling in higher systems. The 14-3-3 protein interact indirectly or directly with many Rho regulators, particularly Rac1 binds directly with 14-3-3. The 14-3-3 proteins also control the activity of the ubiquitous F-actin depolymerizing and severing factor cofilin via binding and stabilizing cellular phosphocofilin levels. Interaction of 14-3-3 with other proteins of the ADF/cofilin family (Twinfilin, Coactosin) warrants further investigation. The binding of 14-3-3 with structural proteins (filamin, clathrin), anchoring proteins (spectrin), sequestering proteins (profilin) have been depicted in the proteome studies.