INIBIOLP   05426
INSTITUTO DE INVESTIGACIONES BIOQUIMICAS DE LA PLATA "PROF. DR. RODOLFO R. BRENNER"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Study of enterocyte FABPs? protein-protein interactions in Caco-2 cells
Autor/es:
B. CÓRSICO; BOTTASSO ARIAS; RODRIGUEZ SAWICKI L
Reunión:
Congreso; Reunion anual Sociedad Arg. Biofisica; 2018
Resumen:
Two isoforms of Fatty Acid Binding Proteins (FABPs) are abundantly expressedwithin intestinal epithelial cells: FABP1 and FABP2. Together they represent 1-2% oftotal proteins present within the enterocytes. Both proteins are associated withintracellular dietary lipid transport and trafficking towards diverse cell fates,although their specific functions are not well understood and are thought to differ.Using the Caco-2 enterocyte cell model, we undertook protein-protein interactionstudies to determine FABP1 and FABP2 protein partners.Firstly, to evaluate FABP1 interactions with other proteins present in Caco-2 celllysate we applied the Far Western blot technique followed by MS analysis. Weidentified two candidate proteins: Heat Shock Protein 60 and calreticulin. On theother side, Co-immunoprecipitation (Co-IP) of FABP1 followed by MS showedinteraction with Creatin kinase B.Secondly, FABP2-protein interactions were analyzed by CoIP. We tested theinteraction between FABP2 and a member of the PPAR (Peroxisome ProliferatorActivated Receptors) transcription factors family, PPARɣ.Thirdly, we examined PPARɣ expression levels in an FABP1 ablated Caco-2 cell linegenerated in our lab applying an antisense technique. The results showed aconcomitant decrease in PPARɣ levels in FABP1 knock down (kd) cells relative tocontrols as well as some PPARs downstream genes.In FABP1 kd cells and controls we performed a luciferase reporter assay with aPPAR response element in frame with the luciferase gene. When FABP1 kd cellswere treated with oleic acid we observed no increase in luciferase responsecompared to the vehicle treatment, opposed to an increase in control cells.In summary, our work shows for the first time the interaction of FABPs withdifferent proteins expressed in Caco-2 enterocytes. Particularly, interaction withPPARs may be indicative of the role of enterocyte FABPs mediating geneexpression. We will conduct further experiments to test this hypothesis.