Three-Dimensional Structure of Bovine Heart Fatty-acid-binding Protein with Bound Palmitic Acid, Determined by Multidimensional NMR Spectroscopy

Lassen, Dirck; Lücke, Christian; Kveder, Marina; Mesgarzadeh, Azita; Schmidt, Joachim W.; Specht, Bernfried; Lezius, Axel G.; Spener, Friedrich; Rüterjans, Heinz

doi:10.1111/j.1432-1033.1995.0266i.x

Cited by 44 publications

(54 citation statements)

References 45 publications

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“…In NMR structural studies of CRABPI (48), the portal elements of apo-forms generally had fewer nuclear Overhauser effect constraints than that for the holo-form. Similar results were obtained during NMR studies of HFABP (50). The largest conformational differences between the crystal and solution structures of HFABP occurred at the ␤-turns between strands ␤E and ␤F, between strands ␤G and ␤H, and in the region of helix ␣II (50).…”

Section: Discussionsupporting

confidence: 74%

The Crystal Structure of the Liver Fatty Acid-binding Protein

Thompson

Winter

Terwey

et al. 1997

Journal of Biological Chemistry

253

274

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The crystal structure of the recombinant form of rat liver fatty acid-binding protein was completed to 2.3 Å and refined to an R factor of 19.0%. The structural solution was obtained by molecular replacement using superimposed polyalanine coordinates of six intracellular lipid-binding proteins as a search probe. The entire amino acid sequence of rat liver fatty acid-binding protein along with an amino-terminal formyl-methionine was modeled in the crystal structure. In addition, the crystal was obtained in the presence of oleic acid, and the initial electron density clearly showed two fatty acid molecules bound within a central cavity. The carboxylate of one fatty acid molecule interacts with arginine 122 and is shielded from free solvent. It has an overall bent conformation. The more solvent-exposed carboxylate of the other oleate is located near the helix-turnhelix that caps one end of the ␤-barrel, while the acyl chain lies in the interior. The cavity contains both polar and nonpolar residues but also shows extensive hydrophobic character around the nonpolar atoms of the ligands. The primary and secondary oleate binding sites appear to be totally interdependent, mainly because favorable hydrophobic interactions form between both aliphatic chains.

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Section: Discussionsupporting

confidence: 74%

The Crystal Structure of the Liver Fatty Acid-binding Protein

Thompson

Winter

Terwey

et al. 1997

Journal of Biological Chemistry

253

274

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“…15 N-labeled bovine heart FABP was an interleaved manner. The water magnetization was oriented expressed using the expression system Escherichia coli either prallel or antiparallel to the magnetization of all other 1 H BL21(DE3)pLysS as described previously [17]. Using a 2-l ferresonances prior to the NOE or ROE mixing time.…”

Section: Methodsmentioning

confidence: 99%

“…Isolation and purification of the protein was corresponding amide protons. performed as described by Lassen and coworkers [17]. RecomThe two-dimensional H 2O-ROE/NOE-1 H, 15 N-HSQC experibinant apo and holo bovine heart FABP were dissolved to a ment [19] on a 13 C/ 15 N-labeled protein sample were aquired with concentration of 1.5 mM in 0.1 M potassium phosphate pH 6.0 32 scans/t 1 increment.…”

Section: Methodsmentioning

confidence: 99%

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Bound water in apo and holo bovine heart fatty‐acid‐binding protein determined by heteronuclear NMR spectroscopy

Mesgarzadeh¹,

Pfeiffer²,

Engelke³

et al. 1998

European Journal of Biochemistry

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Two-and three-dimensional heteronuclear NMR experiments have been performed to identify internally bound water molecules in the solution structure of bovine heart fatty-acid-binding protein (heart FABP). NOE and rotating-frame NOE (ROE) cross peaks between protein protons and protons of bound water molecules were observed in two-dimensional H 2O-ROE/NOE-1 H, 15 N-heteronuclear single quantum coherence spectra recorded from a uniformly 13 C/ 15 N-enriched sample of bovine heart FABP. Contacts between water protons and 23 NH protons of the protein backbone were identified. The protein structure consists of 10 antiparallel β-strands (βAϪβJ), forming two nearly orthogonal β-sheets, and a short helixturn-helix motif connecting β-strands A and B. The spatial folding resembles a β-barrel. Most of the water molecules are localized in the gap between β-strands D and E, and near the two A-helices. In the delipidated heart FABP additional contacts between water molecules and NH protons could be observed using a three-dimensional rotating frame Overhauser 1 H, 15 N heteronuclear single quantum coherence experiment obtained with a 15 N-labeled sample of apo-heart FABP.Keywords : bound water; apoprotein ; holoprotein; fatty-acid-binding protein; NMR.Fatty-acid-binding proteins (FABPs) belong to a class of cy-large cavity. Although the conformations of bound ligands differ tosolic, non-enzymic proteins with high non-covalent affinity for in the distinct FABP types, the conformation of the apo form of long-chain fatty acids and molecular masses of 14Ϫ15 kDa [1, the protein is very similar to the holo form [14]. 2]. FABPs have been isolated from a number of fatty-acid-me-A solution structure of bovine heart FABP has been pretabolizing tissues of mammals, fish, chicken, and insect [2]. It sented previously [15Ϫ17]. The fatty acid ligand is bound in a is proposed that fatty-acid-binding proteins serve as a pool for U-shaped conformation, within the binding cavity [17]. The solubilized fatty acids and protect the cells from the detergent crystal structure of the homologous heart FABP of human museffects of high fatty acid concentrations [3]. They are involved cle with bound fatty acid revealed 13 water molecules within in the cellular uptake of fatty acids and in the intracellular trans-this cavity. Two of these water molecules interact with the carport of fatty acids to the mitochondria. They may modulate en-boxylate group of the fatty acid. A mechanism for fatty acid zyme activities and have an influence on cell growth and cell binding is proposed by Young and coworkers [5] including the differentiation [2]. Nevertheless the precise physiological func-rearrangement of some sidechains and water molecules. Calculation of FABP still remains to be elucidated.tions of the solvent-accessible surface of human muscle heart At present seven distinct FABP types are known which have FABP suggest that the internal cavity is connected to the exterbeen isolated from liver, intestine, heart, adipocyte, myelin-type nal solvent by a portal area w...

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“…Since colony formation in soft agar by (Lassen et al, 1995). The peptide region, which is encoded by the 4th exon, is depicted in black.…”

mentioning

confidence: 99%

Breast cancer growth inhibition by delivery of the MDGI-derived peptide P108

Wang

Kurtz

2000

Oncogene

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Mammary derived growth inhibitor (MDGI) is a member of the family of cytoplasmic fatty acid binding proteins (FABPs), which bind hydrophobic ligands such as fatty acids, retinoids, eicosanoids and prostaglandines. MDGI and an 11 amino acid MDGI-derived conserved Cterminal peptide (P108) inhibits growth of normal mammary epithelial cells in tissue and organ culture, but fails to inhibit proliferation of many breast cancer cell lines in vitro. Here, the e ects of peptide P108 on tumor growth of MCF-7, MDA-MB468 and MDA-MB231 human breast cancer cell lines in nude mice were tested. To deliver P108 into tumors, a novel peptide production system was applied for expression and secretion of small bioactive peptides in mammalian cells. Functional di erentiation was observed in MCF-7 and MDA-MB468 cells upon P108 expression. In addition, EGF-dependent colony formation in soft agar by MDA-MB468 cells was inhibited by secreted P108. Tumor growth in athymic nude mice was suppressed in all three cell lines tested. Furthermore, P108 expressed by MCF-7/P108 cells caused paracrine tumor growth inhibition of MDA-MB231 cells. These results indicate that breast cancer inhibition by P108 is independent of binding to hydrophobic ligands and is perhaps mediated by interference with EGF-dependent signaling pathways.

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Three-Dimensional Structure of Bovine Heart Fatty-acid-binding Protein with Bound Palmitic Acid, Determined by Multidimensional NMR Spectroscopy

Cited by 44 publications

References 45 publications

The Crystal Structure of the Liver Fatty Acid-binding Protein

The Crystal Structure of the Liver Fatty Acid-binding Protein

Bound water in apo and holo bovine heart fatty‐acid‐binding protein determined by heteronuclear NMR spectroscopy

Breast cancer growth inhibition by delivery of the MDGI-derived peptide P108

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