title: I-FABP expression alters the intracellular distribution of the BODIPY C16 fatty acid analog.
abstract: To investigate the structure-function relationships of intestinal fatty acid-binding protein (I-FABP) in cellular fatty acid (FA) trafficking, we compared the distribution of a fluorescent FA analog (BODIPY FL C16) in Cos-1 cells transiently transfected with the wild type protein (wt I-FABP) to that of a variant deleted of the alpha helical domain (HL I-FABP). In vector-only cells, BODIPY fluorescence was distributed throughout the cytoplasm. In the absence of added FA, wt I-FABP was found largely in the perinuclear region with some cytoplasmic staining as well. Addition of BODIPY FL C16 to transfected cells showed that the fluorescent FA was essentially completely colocalized with the protein in the cytoplasmic and perinuclear regions as well as in cytoplasmic clusters that are not observed in the absence of wt I-FABP. For HL I-FABP, the distribution of the protein in the absence of FA was diffusely cytoplasmic, in marked contrast to the wt protein. Addition of BODIPY led to less extensive colocalization than that observed for wt I-FABP. In particular, no localization to the perinuclear region was found. Organelle colocalization studies showed that both proteins colocalized with mitochondria and endoplasmic reticulum/golgi markers, but little with a lysosomal marker. The perinuclear localization for wt I-FABP and BODIPY did not show colocalization with any of the markers tested. Taken together, these results indicate that I-FABP binds FA in vivo and that the helical domain may be important for targeting I-FABP to a perinuclear domain but not, perhaps, to the endoplasmic reticulum, golgi apparatus or mitochondria.
authors.affiliation: INSERM, U476 Nutrition Humaine et Lipides, Marseille, 13385, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Karsenty
authors.forename: Olfa
authors.initials: O
authors.lastname: Helal
authors.forename: Paulette Lechène
authors.initials: PL
authors.lastname: de la Porte
authors.forename: Paule
authors.initials: P
authors.lastname: Beauclair-Deprez
authors.forename: Claire
authors.initials: C
authors.lastname: Martin-Elyazidi
authors.forename: Richard
authors.initials: R
authors.lastname: Planells
authors.forename: Judith
authors.initials: J
authors.lastname: Storch
authors.forename: Marguerite
authors.initials: M
authors.lastname: Gastaldi
mesh_terms: D000818:Animals; D001665:Binding Sites; D001896:Boron Compounds / Q000032:analysis* / Q000378:metabolism; D019556:COS Cells; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000032:analysis*; D008856:Microscopy, Fluorescence; D010169:Palmitic Acids / Q000032:analysis* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: C543305:4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-hexadecanoic acid; D001896:Boron Compounds; D018076:DNA, Complementary; D050556:Fatty Acid-Binding Proteins; D005227:Fatty Acids; D005456:Fluorescent Dyes; D010169:Palmitic Acids