Recombinant liver fatty acid binding protein interacts with fatty acyl-coenzyme A.
AuthorsHubbell T, Behnke WD, Woodford JK, Schroeder F
JournalBiochemistry
PubMed ID8136369
'Rat liver fatty acid binding protein (L-FABP) and rat intestine fatty acid binding protein (I-FABP) are homologous proteins which are both found in intestinal epithelial cells. It was once well accepted that liver fatty acid binding protein bound fatty acyl-CoAs, but the recent finding of a novel acyl-CoA binding protein ... More
Binding of 13-HODE and 15-HETE to phospholipid bilayers, albumin, and intracellular fatty acid binding proteins. implications for transmembrane and intracellular transport and for protection from lipid peroxidation.
AuthorsEk-Von Mentzer BA, Zhang F, Hamilton JA
JournalJ Biol Chem
PubMed ID11278949
'Transport and utilization of fatty acids (FA) in cells is a multistep process that includes adsorption to and movement across the plasma membrane and binding to intracellular fatty acid binding proteins (FABP) in the cytosol. We monitored the transbilayer movement of several polyunsaturated FA and oxidation products (13-hydroxy octadecadienoic acid ... More
Refinement of the structure of Escherichia coli-derived rat intestinal fatty acid binding protein with bound oleate to 1.75-A resolution. Correlation with the structures of the apoprotein and the protein with bound palmitate.
AuthorsSacchettini JC, Scapin G, Gopaul D, Gordon JI
JournalJ Biol Chem
PubMed ID1429698
'The structure of rat intestinal fatty acid binding protein (I-FABP) with bound oleate (C18:1) has been refined with x-ray diffraction data to a resolution of 1.75 A. The protein contains 10 anti-parallel beta strands composed of 99 residues and 2 short helices of 14 residues. Oleate is located in the ... More
Characterization of the sources of protein-ligand affinity: 1-sulfonato-8-(1')anilinonaphthalene binding to intestinal fatty acid binding protein.
AuthorsKirk WR, Kurian E, Prendergast FG
JournalBiophys J
PubMed ID8770188
'1-Sulfonato-8-(1'')anilinonaphthalene (1,8-ANS) was employed as a fluorescent probe of the fatty acid binding site of recombinant rat intestinal fatty acid binding protein (1-FABP). The enhancement of fluorescence upon binding allowed direct determination of binding affinity by fluorescence titration experiments, and measurement of the effects on that affinity of temperature, pH, ... More
Thermodynamics of fatty acid binding to engineered mutants of the adipocyte and intestinal fatty acid-binding proteins.
AuthorsRichieri GV, Low PJ, Ogata RT, Kleinfeld AM
JournalJ Biol Chem
PubMed ID9516437
'We constructed 18 single amino acid mutants of the adipocyte fatty acid-binding protein (A-FABP) and 17 of the intestinal fatty acid-binding protein (I-FABP), at locations in the fatty acid (FA) binding sites. For each mutant protein, we measured thermodynamic parameters that characterize FA binding. Binding affinities ranged from about 200-fold ... More
Fatty acid transfer from liver and intestinal fatty acid-binding proteins to membranes occurs by different mechanisms.
AuthorsHsu KT, Storch J
JournalJ Biol Chem
PubMed ID8662836
'Intestinal absorptive cells contain high levels of expression of two homologous fatty acid-binding proteins (FABP), liver FABP (L-FABP), and intestinal FABP (I-FABP). Both bind long chain fatty acids with relatively high affinity. The functional distinction, if any, between these two proteins remains unknown. It is often hypothesized that FABP are ... More
Binding kinetics of engineered mutants provide insight about the pathway for entering and exiting the intestinal fatty acid binding protein.
AuthorsRichieri GV, Low PJ, Ogata RT, Kleinfeld AM
JournalBiochemistry
PubMed ID10231541
'To better understand the mechanism by which fatty acids bind to and dissociate from the binding cavities of fatty acid binding proteins (FABPs), we constructed 31 single amino acid mutants of the intestinal FABP (I-FABP) and determined the rate constants for binding and dissociation, primarily for long-chain fatty acids (FA). ... More
Interaction of fatty acids with recombinant rat intestinal and liver fatty acid-binding proteins.
AuthorsNemecz G, Hubbell T, Jefferson JR, Lowe JB, Schroeder F
JournalArch Biochem Biophys
PubMed ID1897956
'Intestinal enterocytes contain two homologous fatty acid-binding proteins, intestinal fatty acid-binding protein (I-FABP)2 and liver fatty acid-binding protein (L-FABP). Since the functional basis for this multiplicity is not known, the fatty acid-binding specificity of recombinant forms of both rat I-FABP and rat L-FABP was examined. A systematic comparative analysis of ... More
Fatty acid interactions with a helix-less variant of intestinal fatty acid-binding protein.
AuthorsCistola DP, Kim K, Rogl H, Frieden C
JournalBiochemistry
PubMed ID8652536
'Intestinal fatty acid-binding protein (I-FABP) binds a single molecule of long-chain fatty acid in an enclosed cavity surrounded by two antiparallel beta-sheets. The structure also contains two short alpha-helices which form a cap over one end of the binding cavity adjacent to the methyl terminus of the fatty acid. In ... More
Affinity of fatty acid for (r)rat intestinal fatty acid binding protein:further examination.
AuthorsKurian E, Kirk WR, Prendergast FG
JournalBiochemistry
PubMed ID8620011
'The enhancement of the fluorescence quantum yield of 1,8-anilinonaphthalenesulfonic acid (ANS) upon binding to intestinal fatty acid protein (I-FABP) was exploited to devise an assay for free I-FABP. With this assay, we monitored the competition for free I-FABP between ANS and fatty acids and thereby extracted values for the dissociation ... More
Rat intestinal fatty acid binding protein. A model system for analyzing the forces that can bind fatty acids to proteins.
AuthorsSacchettini JC, Gordon JI
JournalJ Biol Chem
PubMed ID8360139
Intestinal and liver fatty acid binding proteins differentially affect fatty acid uptake and esterification in L-cells.
AuthorsProws DR, Murphy EJ, Schroeder F
JournalLipids
PubMed ID8538377
Differential effects of intestinal (I-FABP) or liver (L-FABP) fatty acid binding proteins on fatty acid uptake and esterification were examined using transfected mouse L-cell fibroblasts. L-FABP, but not I-FABP, expression increased the initial rate and extent of cis-parinaric acid uptake by 50 and 29%, respectively, compared to control cells. I-FABP ... More
Hierarchical folding of intestinal fatty acid binding protein.
AuthorsYeh SR, Ropson IJ, Rousseau DL
JournalBiochemistry
PubMed ID11284675
Intestinal fatty acid binding protein (IFABP) is a member of the lipid binding protein family, members of which have a clam shell type of motif formed by two five-stranded beta-sheets. Understanding the folding mechanism of these proteins has been hindered by the presence of an unresolved burst phase. By initiating ... More
Sterol carrier protein-2, a new fatty acyl coenzyme A-binding protein.
AuthorsFrolov A, Cho TH, Billheimer JT, Schroeder F
JournalJ Biol Chem
PubMed ID8943231
The ability of sterol carrier protein-2 (SCP-2) to interact with long chain fatty acyl-CoAs was examined. SCP-2 bound fluorescent fatty acyl-CoAs at a single site with high affinity. Kd values for cis- and trans-parinaroyl-CoA were 4.5 and 2.8 nM, respectively. Saturated 10-18-carbon and unsaturated 14-20-carbon fatty acyl-CoAs displaced SCP-2-bound fluorescent ... More
Diversity of fatty acid-binding protein structure and function: studies with fluorescent ligands.
AuthorsStorch J
JournalMol Cell Biochem
PubMed ID8232268
The mammalian fatty acid-binding proteins (FABP) are localized in many distinct cell types. They bind long chain fatty acids in vitro, however, their functions and mechanisms of action in vivo remain unknown. The present studies have sought to understand the relationships among these proteins, and to address the possible role ... More
A fluorescently labeled intestinal fatty acid binding protein. Interactions with fatty acids and its use in monitoring free fatty acids.
AuthorsRichieri GV, Ogata RT, Kleinfeld AM
JournalJ Biol Chem
PubMed ID1429693
The fatty acid-binding protein from rat intestine (I-FABP) has been covalently modified with the fluorescent compound Acrylodan. Acrylodan was found to label Lys27, one of the few amino acid residues found by x-ray diffraction studies to change orientation upon fatty acid (FA) binding to I-FABP. Binding of FA to this ... More
Selective binding of cholesterol by recombinant fatty acid binding proteins.
AuthorsNemecz G, Schroeder F
JournalJ Biol Chem
PubMed ID1894612
The sterol binding specificity of rat recombinant liver fatty acid binding protein (L-FABP) and intestinal fatty acid binding protein (I-FABP) was characterized with [3H]cholesterol and a fluorescent sterol analog dehydroergosterol. Ligand binding analysis, fluorescence spectroscopy, and activation of microsomal acyl-CoA:cholesterol acyltransferase activity showed that L-FABP-bound sterols. 1) Lipidex-1000 assay showed ... More
Kinetics of fatty acid interactions with fatty acid binding proteins from adipocyte, heart, and intestine.
AuthorsRichieri GV, Ogata RT, Kleinfeld AM
JournalJ Biol Chem
PubMed ID8626681
Rate constants for the interaction of fatty acids (FA) with fatty acid binding proteins (FABP) from adipocyte (A-FABP), heart (H-FABP), and intestine (I-FABP) were determined by using stopped-flow fluorometry and ADIFAB, the fluorescent probe of free fatty acids (FFA), or a new FFA probe, ADIFAB2, constructed by derivatizing with acrylodan ... More
Escherichia coli-derived rat intestinal fatty acid binding protein with bound myristate at 1.5 A resolution and I-FABPArg106-->Gln with bound oleate at 1.74 A resolution.
AuthorsEads J, Sacchettini JC, Kromminga A, Gordon JI
JournalJ Biol Chem
PubMed ID8253762
Rat intestinal fatty acid binding protein (I-FABP) is a 131-residue protein composed of two short alpha-helices (alpha I and alpha II) and 10 anti-parallel beta-strands organized into two nearly orthogonal beta-sheets. The structure of crystalline I-FABP with bound tetradecanoate (myristate) has been refined to a resolution of 1.5 A and ... More
Thermodynamics of fatty acid binding to fatty acid-binding proteins and fatty acid partition between water and membranes measured using the fluorescent probe ADIFAB.
AuthorsRichieri GV, Ogata RT, Kleinfeld AM
JournalJ Biol Chem
PubMed ID7797491
Using the fluorescent probe ADIFAB (acrylodan-derivatized intestinal fatty acid-binding protein) to determine the equilibrium concentration of the free (unbound) fatty acid (FFA), dissociation constants were measured between 10 and 50 degrees C for the interaction of five different long chain fatty acids (FA) with fatty acid-binding proteins (FABP) from adipocyte, ... More
Polyene fatty acid interactions with recombinant intestinal and liver fatty acid-binding proteins. Spectroscopic studies.
AuthorsNemecz G, Jefferson JR, Schroeder F
JournalJ Biol Chem
PubMed ID1894608
Binding and proximity relationships of fatty acids with recombinant rat liver fatty acid-binding protein (L-FABP) and intestinal fatty acid-binding protein (I-FABP) were studied with absorption and fluorescence spectroscopy. Protein aromatic amino acids were examined in the absence and presence of bound fatty acid. Second derivative absorbance spectroscopy of the apo- ... More
Site-directed mutagenesis was used to introduce cysteine residues into the rat intestinal fatty acid binding protein, an almost all beta-sheet protein that in the wild-type contains neither cysteine nor proline residues. Six mutants (I23C, S53C, V60C, L72C, L89C, and A104C) with a single cysteine residue substituted for a hydrophobic residue ... More
Intestinal fatty acid-binding protein: the structure and stability of a helix-less variant.
AuthorsKim K, Cistola DP, Frieden C
JournalBiochemistry
PubMed ID8652535
The structure of Escherichia coli-derived rat intestinal fatty acid-binding protein (I-FABP) exhibits a beta-clam topology comprised of two five-stranded antiparallel beta-sheets surrounding a large solvent-filled cavity into which the ligand binds. It also contains two alpha-helices that span residues E15-A32 and join beta-strands A and B. This helical domain is ... More