PDBsum entry 1acd

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Fatty acid binding protein PDB id
Protein chain
131 a.a. *
* Residue conservation analysis
PDB id:
Name: Fatty acid binding protein
Title: V32d/f57h mutant of murine adipocyte lipid binding protein
Structure: Adipocyte lipid binding protein. Chain: a. Synonym: albp. Engineered: yes. Mutation: yes
Source: Mus musculus. House mouse. Organism_taxid: 10090. Cell: adipocyte. Expressed in: escherichia coli. Expression_system_taxid: 562
2.70Å     R-factor:   0.198     R-free:   0.276
Authors: J.Ory,C.D.Kane,M.Simpson,L.J.Banaszak,D.A.Bernlohr
Key ref:
J.Ory et al. (1997). Biochemical and crystallographic analyses of a portal mutant of the adipocyte lipid-binding protein. J Biol Chem, 272, 9793-9801. PubMed id: 9092513 DOI: 10.1074/jbc.272.15.9793
06-Feb-97     Release date:   16-Jun-97    
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Protein chain
Pfam   ArchSchema ?
P04117  (FABP4_MOUSE) -  Fatty acid-binding protein, adipocyte
132 a.a.
131 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 4 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   3 terms 
  Biological process     cellular response to lithium ion   9 terms 
  Biochemical function     transporter activity     2 terms  


DOI no: 10.1074/jbc.272.15.9793 J Biol Chem 272:9793-9801 (1997)
PubMed id: 9092513  
Biochemical and crystallographic analyses of a portal mutant of the adipocyte lipid-binding protein.
J.Ory, C.D.Kane, M.A.Simpson, L.J.Banaszak, D.A.Bernlohr.
A number of crystallographic studies of the adipocyte lipid-binding protein have established that the fatty acid-binding site is within an internalized water-filled cavity. The same studies have also suggested the existence of a region physically distinct from the fatty acid-binding site which connects the cavity of the protein with the external solvent, hereafter referred to as the portal. In an effort to examine the portal region, we have used site-directed mutagenesis to introduce the mutations V32D/F57H into the murine ALBP cDNA. Mutant protein has been isolated, crystallized, and its stability and binding properties studied by biochemical methods. As assessed by guanidine-HCl denaturation, the mutant form exhibited a slight overall destabilization relative to the wild-type protein under both acid and alkaline conditions. Accessibility to the cavity in both the mutant and wild-type proteins was observed by stopped-flow analysis of the modification of a cavity residue, Cys117, by the sulfhydryl reactive agent 5, 5'-dithiobis(2-nitrobenzoic acid) at pH 8.5. Cys117 of V32D/F57H ALBP was modified 7-fold faster than the wild-type protein. The ligand binding properties of both the V32D/F57H mutant and wild-type proteins were analyzed using a fluorescent probe at pH 6.0 and 8.0. The apparent dissociation constants for 1-anilinonaphthalene-8-sulfonic acid were approximately 9-10-fold greater than the wild-type protein, independent of pH. In addition, there is a 6-fold increase in the Kd for oleic acid for the portal mutant relative to the wild-type at pH 8.0. To study the effect of pH on the double mutant, it was crystallized and analyzed in two distinct space groups at pH 4.5 and 6.4. While in general the differences in the overall main chain conformations are negligible, changes were observed in the crystallographic structures near the site of the mutations. At both pH values, the mutant side chains are positioned somewhat differently than in wild-type protein. To ensure that the mutations had not altered ionic conditions near the binding site, the crystallographic coordinates were used to monitor the electrostatic potentials from the head group site to the positions near the portal region. The differences in the electrostatic potentials were small in all regions, and did not explain the differences in ligand affinity. We present these results within the context of fatty acid binding and suggest lipid association is more complex than that described within a single equilibrium event.
  Selected figure(s)  
Figure 4.
Fig. 4. The crystal structures of the V32D/F57H mutant near the portal region. The stereodiagram contains a ball and stick representation of the crystal structures of the V32D/F57H mutant near the expected portal region of ALBP. Notice His57 is hydrogen bonded to Ser55 in the pH 4.5 form, but makes a bond with Asp32 in the pH 6.4 form. The methyl end of HDS is shown as well, illustrating the residues close proximity to the ligand. HDS binding was modeled^ after the V32D/F57H mutant coordinates were aligned with the crystal structure of ALBP containing bound HDS.
Figure 6.
Fig. 6. Schematic representation of the open and closed conformations of ALBP. Schematic representation of the apo- and holoprotein forms of ALBP in their open and closed conformations. Alternation of the ligand conformation between the open and closed conformations is speculative.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (1997, 272, 9793-9801) copyright 1997.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19834748 L.B.Levin, A.Ganoth, S.Amram, E.Nachliel, M.Gutman, and Y.Tsfadia (2010).
Insight into the interaction sites between fatty acid binding proteins and their ligands.
  J Mol Model, 16, 929-938.  
19564911 D.Long, Y.Mu, and D.Yang (2009).
Molecular dynamics simulation of ligand dissociation from liver fatty acid binding protein.
  PLoS One, 4, e6081.  
19118410 L.B.Levin, E.Nachliel, M.Gutman, and Y.Tsfadia (2009).
Molecular dynamics study of the interaction between fatty acid binding proteins with palmitate mini-micelles.
  Mol Cell Biochem, 326, 29-33.  
19595785 Q.Wang, T.Guan, H.Li, and D.A.Bernlohr (2009).
A novel polymorphism in the chicken adipocyte fatty acid-binding protein gene (FABP4) that alters ligand-binding and correlates with fatness.
  Comp Biochem Physiol B Biochem Mol Biol, 154, 298-302.  
17660261 M.Mihajlovic, and T.Lazaridis (2007).
Modeling fatty acid delivery from intestinal fatty acid binding protein to a membrane.
  Protein Sci, 16, 2042-2055.  
17761196 R.E.Gillilan, S.D.Ayers, and N.Noy (2007).
Structural basis for activation of fatty acid-binding protein 4.
  J Mol Biol, 372, 1246-1260.
PDB codes: 2q9s 2qm9
16361342 R.Friedman, E.Nachliel, and M.Gutman (2006).
Fatty acid binding proteins: same structure but different binding mechanisms? Molecular dynamics simulations of intestinal fatty acid binding protein.
  Biophys J, 90, 1535-1545.  
10423455 J.J.Ory, and L.J.Banaszak (1999).
Studies of the ligand binding reaction of adipocyte lipid binding protein using the fluorescent probe 1, 8-anilinonaphthalene-8-sulfonate.
  Biophys J, 77, 1107-1116.
PDB code: 2ans
10026291 P.Penzes, and J.L.Napoli (1999).
Holo-cellular retinol-binding protein: distinction of ligand-binding affinity from efficiency as substrate in retinal biosynthesis.
  Biochemistry, 38, 2088-2093.  
9555061 N.R.Coe, and D.A.Bernlohr (1998).
Physiological properties and functions of intracellular fatty acid-binding proteins.
  Biochim Biophys Acta, 1391, 287-306.  
9849941 V.J.LiCata, and D.A.Bernlohr (1998).
Surface properties of adipocyte lipid-binding protein: Response to lipid binding, and comparison with homologous proteins.
  Proteins, 33, 577-589.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.