We expect to identify the interaction partners of intestinal FABPs in order to describe the molecular crosstalk relevant for its biological functions.Facultad de Ciencias Médica

Córsico, Betina

Falomir Lockhart, Lisandro J.

Franchini, Gisela Raquel

Guerbi, María Ximena

Storch, Judith

El Servicio de Difusión de la Creación Intelectual

 
 
 
 
2010 Octubre, 2(1): 1- 
 
1 
 
UNRAVELING INTESTINAL FATTY ACID BINDING PROTEINS FUNCTIONS: ANALYSIS 
OF FABP-MEMBRANE AND FABP-PROTEIN INTERACTIONS 
 
Autores 
Lisandro J. Falomir Lockhart#+, María Ximena Guerbi#, Gisela R. Franchinni#, Judith Storch¶ and Betina 
Córsico#+. 
Lugar de Trabajo 
#  INIBIOLP, University of La Plata, Calle 60 y120, 1900, La Plata, Argentina. 
+ Consejo Nacional de Investigaciones Científicas y Técnicas, Avda. Rivadavia 1917, C1033AAJ, Cdad. de 
Buenos Aires, Argentina. 
¶ Department of Nutritional Sciences, Rutgers the State University of New Jersey, 96 Lipman Dr, New 
Brunswick, NJ, 08901-8525, U.S.A. 
e-mail de contacto (IMPORTANTE): lysfalomir@yahoo.com.ar, bcorsico@atlas.med.unlp.edu.ar 
 
Introducción 
Intestinal enterocytes express large concentrations of two homologous fatty acid binding 
proteins (FABPs): intestinal FABP (IFABP) (15.1 kDa) and liver FABP (LFABP) (14.2 kDa). It 
has long been hypothesized that FABPs participate in the intracellular transport and 
processing of the large quantities of fatty acids (FA) absorbed by the intestine, but their 
specific function are still poorly understood. In particular, it is currently not known why a single 
cell type contains two distinct types of FABP. 
 
Objetivos 
We expect to identify the interaction partners of intestinal FABPs in order to describe the 
molecular crosstalk relevant for its biological functions. 
 
Materiales y métodos 
In this work, we took advantage of different structural variants of intestinal FABPs and a 
battery of biochemical and biophysical methodologies to analyze its interaction with 
membranes and other proteins. Tb/DPA complex leakage, binding to sucrose loaded vesicles, 
Cyt c competition assay and radiolabeled photoactivable phospholipid crosslinking were 
employed to study different factors (vesicle curvature, ligand nature, lipid composition, etc) 
that modulate the interaction of FABPs with artificial model membranes. On the other hand, a 
fluorescence based assay was employed to analyze colisional transfer of fatty acid analogs 
between IFABP and LFABP in a stopped-flow module. 
 
Resultados 
Lipid composition and ligand binding show strong changes in membrane interaction 
properties of IFABP and LFABP, as evidenced by different techniques. Collisional transfer of 
fluorescent fatty acids suggests functional interaction between these intestinal FABPs. 
 
Conclusiones 
Intestinal FABPs could interact differentially with each other and with subcellular membranous 
fractions accordingly with ligand binding. The results in vitro presented here may reflect in 
vivo functions of intestinal FABPs. Increasing evidence supports the idea that FABPs 
participate and modulate several cell functions and further studies will be needed to unravel 
its physiological role. 
 
 
 
 
 


English

Unraveling intestinal fatty acid binding proteins functions: analysis of FABP-membrane and FABP-protein interactions

Servicio de Difusión de la Creación Intelectual

    2010 Octubre, 2(1): 1-  1  UNRAVELING INTESTINAL FATTY ACID BINDING PROTEINS FUNCTIONS: ANALYSIS OF FABP-MEMBRANE AND FABP-PROTEIN INTERACTIONS  Autores Lisandro J. Falomir Lockhart#+, María Ximena Guerbi#, Gisela R. Franchinni#, Judith Storch¶ and Betina Córsico#+. Lugar de Trabajo #  INIBIOLP, University of La Plata, Calle 60 y120, 1900, La Plata, Argentina. + Consejo Nacional de Investigaciones Científicas y Técnicas, Avda. Rivadavia 1917, C1033AAJ, Cdad. de Buenos Aires, Argentina. ¶ Department of Nutritional Sciences, Rutgers the State University of New Jersey, 96 Lipman Dr, New Brunswick, NJ, 08901-8525, U.S.A. e-mail de contacto (IMPORTANTE): lysfalomir@yahoo.com.ar, bcorsico@atlas.med.unlp.edu.ar  Introducción Intestinal enterocytes express large concentrations of two homologous fatty acid binding proteins (FABPs): intestinal FABP (IFABP) (15.1 kDa) and liver FABP (LFABP) (14.2 kDa). It has long been hypothesized that FABPs participate in the intracellular transport and processing of the large quantities of fatty acids (FA) absorbed by the intestine, but their specific function are still poorly understood. In particular, it is currently not known why a single cell type contains two distinct types of FABP.  Objetivos We expect to identify the interaction partners of intestinal FABPs in order to describe the molecular crosstalk relevant for its biological functions.  Materiales y métodos In this work, we took advantage of different structural variants of intestinal FABPs and a battery of biochemical and biophysical methodologies to analyze its interaction with membranes and other proteins. Tb/DPA complex leakage, binding to sucrose loaded vesicles, Cyt c competition assay and radiolabeled photoactivable phospholipid crosslinking were employed to study different factors (vesicle curvature, ligand nature, lipid composition, etc) that modulate the interaction of FABPs with artificial model membranes. On the other hand, a fluorescence based assay was employed to analyze colisional transfer of fatty acid analogs between IFABP and LFABP in a stopped-flow module.  Resultados Lipid composition and ligand binding show strong changes in membrane interaction properties of IFABP and LFABP, as evidenced by different techniques. Collisional transfer of fluorescent fatty acids suggests functional interaction between these intestinal FABPs.  Conclusiones Intestinal FABPs could interact differentially with each other and with subcellular membranous fractions accordingly with ligand binding. The results in vitro presented here may reflect in vivo functions of intestinal FABPs. Increasing evidence supports the idea that FABPs participate and modulate several cell functions and further studies will be needed to unravel its physiological role.      

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Unraveling intestinal fatty acid binding proteins functions: analysis of FABP-membrane and FABP-protein interactions

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