Characterization of crystalline rat liver fatty acid binding protein produced in Escherichia coli.

Rat liver fatty acid-binding protein is able to accommodate a wide range of non-polar anions in addition to long-chain fatty acids. The two arginine residues of rat liver fatty acid-binding protein, Arg122 and Arg126, have been mutated and the effect of mutation on ligand binding investigated. No significant decrease in affinity for the fluorescent fatty acid analogue, 11-(5-dimethylaminonaphthalenesulphonyl amino)undecanoic acid, or oleate was observed. However, the apparent affinity for oleoyl-CoA was slightly increased with the mutations Ala122 and Gln122 such that oleoyl-CoA rather than oleate became the preferred ligand for these mutants. Small changes in protein stability were observed with the Arg122 mutations. The lack of notable ionic involvement of the conserved internal residue Arg122 in ligand binding is consistent with the hypothesis that the mode of ligand binding in liver fatty acid-binding protein is markedly different from that of other members of this lipid-binding protein family.

Download Full-text

Growth hormone and aging change rat liver fatty acid binding protein levels.

Journal of the American College of Nutrition ◽

10.1080/07315724.1996.10718584 ◽

1996 ◽

Vol 15 (2) ◽

pp. 169-174 ◽

Cited By ~ 8

Author(s):

S S Singer ◽

K Henkels ◽

A Deucher ◽

M Barker ◽

J Singer ◽

...

Keyword(s):

Growth Hormone ◽

Fatty Acid ◽

Rat Liver ◽

Fatty Acid Binding Protein ◽

Binding Protein ◽

Liver Fatty Acid ◽

Fatty Acid Binding ◽

Protein Levels ◽

Acid Binding Protein ◽

Aging Change

Download Full-text

The binding of natural and fluorescent lysophospholipids to wild-type and mutant rat liver fatty acid-binding protein and albumin

Biochemical Journal ◽

10.1042/bj3070305 ◽

1995 ◽

Vol 307 (1) ◽

pp. 305-311 ◽

Cited By ~ 16

Author(s):

A E A Thumser ◽

D C Wilton

Keyword(s):

Fatty Acid ◽

Rat Liver ◽

Fatty Acid Binding Protein ◽

Binding Protein ◽

Liver Fatty Acid ◽

Fatty Acid Binding ◽

Acid Binding Protein ◽

Kd Values ◽

Fluorescence Characteristics ◽

Wide Range

Rat liver fatty acid-binding protein (FABP) is able to bind a wide range of non-polar anionic ligands, including lysophospholipids. In order to understand the nature of lysophospholipid interactions with liver FABP, the binding of natural lysophospholipids and two fluorescent analogues, N-(5-dimethylaminonaphthalenesulphonyl)-1-palmitoyl-sn-glycero-3- phosphoethanolamine (dansyl lysoPE) and 1-(O-[11-(5-dimethylaminonaphthalene-sulphonyl)amino]undecyl)-sn-glycero -3- phosphocholine (dansyl-C11-lysoPAF), has been investigated. The results confirmed the ability of liver FABP to bind lysophospholipids with KD values in the range of 1-2 microM, and a 1:1 binding stoichiometry was indicated. Binding of fluorescent lysophospholipids was enhanced with the FABP mutant, R122Q, possibly due to increased flexibility of the binding cavity as a result of reduced hydrogen-bonding constraints. The fluorescent lysophospholipids also bound to albumin, with KD values in the range 0.1-1.0 microM, and could be displaced by oleic acid. The fluorescence characteristics of the dansyl lysophospholipid analogue dansyl-C11-lyso-PAF suggested that this probe binds to the same site(s) on albumin as the fluorescent fatty acid probe 11-(5-dimethylaminonaphthalene-sulphonylamino)-undecanoic acid (DAUDA).

Download Full-text