caa a22 4500 477136419 CHVBK 20180405111719.0 cr unu---uuuuu 170330e19970401xx s 000 0 eng 10.1023/A:1018666522787 doi (NATIONALLICENCE)springer-10.1023/A:1018666522787 Solution structure of human intestinal fatty acid binding protein: Implications for ligand entry and exit [Elektronische Daten] [Fengli Zhang, Christian Lücke, Leslie Baier, James Sacchettini, James Hamilton] The human intestinal fatty acid binding protein (I-FABP) is a small (131 amino acids) proteinwhich binds dietary long-chain fatty acids in the cytosol of enterocytes. Recently, an alanineto threonine substitution at position 54 in I-FABP has been identified which affects fatty acidbinding and transport, and is associated with the development of insulin resistance in severalpopulations including Mexican-Americans and Pima Indians. To investigate the molecularbasis of the binding properties of I-FABP, the 3D solution structure of the more commonform of human I-FABP (Ala54) was studied by multidimensional NMR spectroscopy.Recombinant I-FABP was expressed from E. coli in the presence and absence of 15N-enriched media. The sequential assignments for non-delipidated I-FABP were completed byusing 2D homonuclear spectra (COSY, TOCSY and NOESY) and 3D heteronuclear spectra(NOESY-HMQC and TOCSY-HMQC). The tertiary structure of human I-FABP wascalculated by using the distance geometry program DIANA based on 2519 distance constraintsobtained from the NMR data. Subsequent energy minimization was carried out by using theprogram SYBYL in the presence of distance constraints. The conformation of human I-FABPconsists of 10 antiparallel β-strands which form two nearly orthogonal β-sheets offive strands each, and two short α-helices that connect the β-strands A and B. Theinterior of the protein consists of a water-filled cavity between the two β-sheets. TheNMR solution structure of human I-FABP is similar to the crystal structure of rat I-FABP.The NMR results show significant conformational variability of certain backbone segmentsaround the postulated portal region for the entry and exit of fatty acid ligand. Kluwer Academic Publishers, 1997 Human intestinal fatty acid binding protein nationallicence Isotope enrichment nationallicence Multidimensional NMR spectroscopy nationallicence Sequential assignments nationallicence Solution structure nationallicence Zhang Fengli Department of Biophysics, Boston University School of Medicine, 80 East Concord Street, 02118, Boston, MA, U.S.A aut Lücke Christian Johann Wolfgang Goethe-Universität, Marie-Curie-Straße 9, D-60439, Frankfurt am Main, Germany aut Baier Leslie Phoenix Epidemiology and Clinical Research Branch, NIDDK, NIH, 85016, Phoenix, AZ, U.S.A aut Sacchettini James Department of Biochemistry and Biophysics, Texas A+M University, 77845, College Station, TX, U.S.A aut Hamilton James Department of Biophysics, Boston University School of Medicine, 80 East Concord Street, 02118, Boston, MA, U.S.A aut Journal of Biomolecular NMR Kluwer Academic Publishers 9/3(1997-04-01), 213-228 0925-2738 9:3<213 1997 9 10858 https://doi.org/10.1023/A:1018666522787 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018666522787 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Fengli Department of Biophysics, Boston University School of Medicine, 80 East Concord Street, 02118, Boston, MA, U.S.A aut NATIONALLICENCE 700 1- Lücke Christian Johann Wolfgang Goethe-Universität, Marie-Curie-Straße 9, D-60439, Frankfurt am Main, Germany aut NATIONALLICENCE 700 1- Baier Leslie Phoenix Epidemiology and Clinical Research Branch, NIDDK, NIH, 85016, Phoenix, AZ, U.S.A aut NATIONALLICENCE 700 1- Sacchettini James Department of Biochemistry and Biophysics, Texas A+M University, 77845, College Station, TX, U.S.A aut NATIONALLICENCE 700 1- Hamilton James Department of Biophysics, Boston University School of Medicine, 80 East Concord Street, 02118, Boston, MA, U.S.A aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 9/3(1997-04-01), 213-228 0925-2738 9:3<213 1997 9 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer