Fatty acid binding proteins (FABP) are a family of small cytosolic proteins that are abundant in a variety of tissues and in many organisms. It is likely that FABPs play important roles in the transport and metabolism of fatty acids. In this event alterations in these proteins might be involved in diseases in which fat metabolism is implicated and indeed recent evidence suggests that modification of the intestinal FABP may be linked to diabetes. Because there is considerable amino acid sequence variation in FABPs from different tissues it has generally been thought that the biochemical properties of these proteins would be tissue specific, yet until recently no evidence existed for such variability. During the past three years, however, our studies have demonstrated that the thermodynamic and kinetic properties of FA-FABP interactions are extremely sensitive both to the tissue origin of the FABP and to the molecular species of FA. The long-term goals of the present studies are (1) to provide a rigorous biochemical basis for understanding the physiologic role of these proteins, (2) to understand the molecular interactions that give rise to the observed tissue-specific heterogeneity of binding, and (3) to develop a variety of fluorescent probes of free fatty acids (FFA) that will have application in clinical medicine as well as biochemistry. Because FABPs can be grouped according to their sequence homologies and our initial studies of three of these groups reveal that thermodynamic features of FA binding are characteristic of these groups, he will first extend these kinds of studies to members of other groups. For each one of these groups we will then use site directed mutagenesis to alter amino acid residues that interact in the binding cavity and determine the interaction energies corresponding to each residue change. Finally, he will use the results of these studies to construct new FFA probes with novel binding and fluorescent spectroscopic properties.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM046931-08
Application #
6018876
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1992-08-01
Project End
2001-07-31
Budget Start
1999-08-01
Budget End
2001-07-31
Support Year
8
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Torrey Pines Institute for Molecular Studies
Department
Type
DUNS #
605758754
City
San Diego
State
CA
Country
United States
Zip Code
Richieri, G V; Ogata, R T; Zimmerman, A W et al. (2000) Fatty acid binding proteins from different tissues show distinct patterns of fatty acid interactions. Biochemistry 39:7197-204
Richieri, G V; Low, P J; Ogata, R T et al. (1999) Binding kinetics of engineered mutants provide insight about the pathway for entering and exiting the intestinal fatty acid binding protein. Biochemistry 38:5888-95
Richieri, G V; Ogata, R T; Kleinfeld, A M (1999) Fatty acid interactions with native and mutant fatty acid binding proteins. Mol Cell Biochem 192:77-85
Richieri, G V; Ogata, R T; Kleinfeld, A M (1999) The measurement of free fatty acid concentration with the fluorescent probe ADIFAB: a practical guide for the use of the ADIFAB probe. Mol Cell Biochem 192:87-94
Richieri, G V; Low, P J; Ogata, R T et al. (1998) Thermodynamics of fatty acid binding to engineered mutants of the adipocyte and intestinal fatty acid-binding proteins. J Biol Chem 273:7397-405
Richieri, G V; Low, P J; Ogata, R T et al. (1997) Mutants of rat intestinal fatty acid-binding protein illustrate the critical role played by enthalpy-entropy compensation in ligand binding. J Biol Chem 272:16737-40
Richieri, G V; Ogata, R T; Kleinfeld, A M (1996) Thermodynamic and kinetic properties of fatty acid interactions with rat liver fatty acid-binding protein. J Biol Chem 271:31068-74
Richieri, G V; Ogata, R T; Kleinfeld, A M (1996) Kinetics of fatty acid interactions with fatty acid binding proteins from adipocyte, heart, and intestine. J Biol Chem 271:11291-300
Richieri, G V; Ogata, R T; Kleinfeld, A M (1995) Thermodynamics of fatty acid binding to fatty acid-binding proteins and fatty acid partition between water and membranes measured using the fluorescent probe ADIFAB. J Biol Chem 270:15076-84
Richieri, G V; Ogata, R T; Kleinfeld, A M (1994) Equilibrium constants for the binding of fatty acids with fatty acid-binding proteins from adipocyte, intestine, heart, and liver measured with the fluorescent probe ADIFAB. J Biol Chem 269:23918-30