Molecular and Cellular Biochemistry

, Volume 192, Issue 1–2, pp 109–121 | Cite as

A comparative study of the backbone dynamics of two closely related lipid binding proteins: Bovine heart fatty acid binding protein and porcine ileal lipid binding protein

  • Christian Lücke
  • David Fushman
  • Christian Ludwig
  • James A. Hamilton
  • James C. Sacchettini
  • Heinz Rüterjans


The backbone dynamics of bovine heart fatty acid binding protein (H-FABP) and porcine ileal lipid binding protein (ILBP) were studied by 15N NMR relaxation (T1 and T2) and steady state heteronuclear 15N{1H} NOE measurements. The microdynamic parameters characterizing the backbone mobility were determined using the ‘model-free’ approach. For H-FABP, the non-terminal backbone amide groups display a rather compact protein structure of low flexibility. In contrast, for ILBP an increased number of backbone amide groups display unusually high internal mobility. Furthermore, the data indicate a higher degree of conformational exchange processes in the μsec-msec time range for ILBP compared to H-FABP. These results suggest significant differences in the conformational stability for these two structurally highly homologous members of the fatty acid binding protein family.

lipid binding protein 15N relaxation protein backbone dynamics model-free approach 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Veerkamp JH, Maatman RGHJ: Cytoplasmic fatty acid-binding proteins: their structure and genes. Prog Lipid Res 34: 17–52, 1995Google Scholar
  2. 2.
    Banaszak L, Winter N, Xu Z, Bernlohr DA, Cowan S, Jones TA: Lipid-binding proteins: A family of fatty acid and retinoid transport proteins. Adv Prot Chem 45: 89–151, 1994Google Scholar
  3. 3.
    Sacchettini JC, Gordon JI, Banaszak LJ: The structure of crystalline Escherichia coli-derived rat intestinal fatty acid-binding protein at 2.5–Å resolution. J Biol Chem 263: 5815–5819, 1988Google Scholar
  4. 4.
    Sacchettini JC, Gordon JI, Banaszak LJ: Refined apoprotein structure of rat intestinal fatty acid binding protein produced in Escherichia coli. Proc Natl Acad Sci USA 86: 7736–7740, 1989Google Scholar
  5. 5.
    Scapin G, Gordon JI, Sacchettini JC: Refinement of the structure of recombinant rat intestinal fatty acid-binding apoprotein at 1.2–Å resolution. J Biol Chem 267: 4253–4269, 1992Google Scholar
  6. 6.
    Scapin G, Spadon P, Mammi M, Zanotti G, Monaco HL: Crystal structure of chicken liver basic fatty acid-binding protein at 2.7 Å resolution. Mol Cell Biochem 98: 95–99, 1990Google Scholar
  7. 7.
    Thompson J, Winter N, Terwey D, Bratt J, Banaszak LJ: The crystal structure of the liver fatty acid-binding protein. A complex with two bound oleates. J Biol Chem 272: 7140–7150, 1997Google Scholar
  8. 8.
    Müller-Fahrnow A, Egner U, Jones TA, Rüdel H, Spener F, Saenger W: Three-dimensional structure of fatty-acid-binding protein from bovine heart. Eur J Biochem 199: 271–276, 1991Google Scholar
  9. 9.
    Zanotti G, Scapin G, Spadon P, Veerkamp JH, Sacchettini JC: Three-dimensional structure of recombinant human muscle fatty acid-binding protein. J Biol Chem 267: 18541–18550, 1992Google Scholar
  10. 10.
    Young ACM, Scapin G, Kromminga A, Patel SB, Veerkamp JH, Sacchettini JC: Structural studies on human muscle fatty acid binding protein at 1.4 Å resolution: Binding interactions with three C18 fatty acids. Structure 2: 523–534, 1994Google Scholar
  11. 11.
    Xu Z, Bernlohr DA, Banaszak LJ: Crystal structure of recombinant murine adipocyte lipid binding protein. Biochemistry 31: 3484–3492, 1992Google Scholar
  12. 12.
    Jones TA, Bergfors T, Sedzik J, Unge T: The three-dimensional structure of P2 myelin protein. EMBO J 7: 1597–1604, 1988Google Scholar
  13. 13.
    Cowan SW, Newcomer ME, Jones TA: Crystallographic studies on a family of cellular lipophilic transport proteins: The refinement of P2 myelin protein and the structure determination and refinement of cellular retinol-binding protein in complex with all-trans retinol. J Mol Biol 230: 1225–1246, 1993Google Scholar
  14. 14.
    Benning MM, Smith AF, Wells MA, Holden HM: Crystallization, structure determination and least-squares refinement to 1.75 Å resolution of the fatty acid binding protein isolated from Manducta sexta L. J Mol Biol 228: 208–219, 1992Google Scholar
  15. 15.
    Winter NS, Bratt JM, Banaszak LJ: Crystal structures of holo and apo-cellular retinol-binding protein II. J Biol Chem 230: 1247–1259, 1993Google Scholar
  16. 16.
    Lassen D, Lücke C, Kveder M, Mesgarzadeh A, Schmidt JM, Specht B, Lezius A, Spener F, Rüterjans H: Three-dimensional structure of bovine heart fatty-acid-binding protein with bound palmitic acid, determined by multidimensional NMR spectroscopy. Eur J Biochem 230: 266–280, 1995Google Scholar
  17. 17.
    Lücke C, Zhang F, Rüterjans H, Hamilton JA, Sacchettini JC: Flexibility is a likely determinant of binding in the case of ileal lipid binding protein. Structure 4: 785–800, 1996Google Scholar
  18. 18.
    Zhang F, Ñcke C, Baier LJ, Sacchettini JC, Hamilton JA: Solution structure of human intestinal fatty acid binding protein: Implications for ligand entry and exit. J Biomol NMR 9: 213–228, 1997Google Scholar
  19. 19.
    Hodsdon ME, Cistola DR: Ligand binding alters the backbone mobility of intestinal fatty acid binding protein as monitored by 15N relaxation and 1H exchange. Biochemistry 36: 2278–2290, 1997Google Scholar
  20. 20.
    Jakoby MG IV, Miller KR, Toner JJ, Bauman A, Cheng L, Li E, Cistola DR: Ligand-protein electrostatic interactions govern the specificity of retinol-and fatty acid-binding proteins. Biochemistry 32: 872–878, 1993Google Scholar
  21. 21.
    Eads J, Sacchettini JC, Kromminga A, Gordon R: Escherichia coli-derived rat intestinal fatty acid binding protein with bound myristate at 1.5 Å resolution and I-FABPArg106–>Gln with bound oleate at 1.74 Å resolution. J Biol Chem 268: 26375–26385, 1993Google Scholar
  22. 22.
    Sacchettini JC, Hauft SM, Van Camp SL, Cistola DP, Gordon JI: Developmental and structural studies of an intracellular lipid binding protein expressed in the ileal epithelium. J Biol Chem 265: 19199–19207, 1990Google Scholar
  23. 23.
    Kay LE, Torchia DA, Bax A: Backbone dynamics of proteins as studied by nitrogen-15 inverse detected heteronuclear NMR spectroscopy: Application to staphylococcal nuclease. Biochemistry 28: 8972–8979, 1989Google Scholar
  24. 24.
    Fushman D, Weisemann R, Thüring H, Rüterjans H: Backbone dynamics of ribonuclease T1 and its complex with 2′GMP studied by two-dimensional heteronuclear NMR spectroscopy. J Biomol NMR 4: 61–78, 1994Google Scholar
  25. 25.
    Gaudin F, Paquet F, Chanteloup L, Beau JM, Nguen TT, Lancelot G: Selectively 13C-enriched DNA: Dynamics of the C1′-H1′ vector in d(CGAAATTTVG)3. J Biomol NMR 5: 49–58, 1995Google Scholar
  26. 26.
    King GC, Harper JW, Xi Z: Isotope labeling for 13C relaxation measurements on RNA. Meth Enzymol 261: 436–50, 1995Google Scholar
  27. 27.
    Shaka AJ, Barker PB, Freeman R: Computer-optimized decoupling scheme for wideband applications and low-level operation. J Magn Res 64: 547–552, 1985Google Scholar
  28. 28.
    Morris GA, Freeman R: Selective excitation in fourier transform nuclear magnetic resonance. J Magn Res 29: 433–462, 1978Google Scholar
  29. 29.
    Kay LE, Nicholson LK, Delaglio F, Bax A, Torchia DA: Pulse sequences for removal of the effects of cross correlation between dipolar and chemical-shift anisotropy relaxation mechanisms on the measurement of heteronuclear T1 and T2 values in proteins. J Magn Res 97: 359–375, 1992Google Scholar
  30. 30.
    Grzesiek S, Bax A: The importance of not saturating H2O in protein NMR. Application to sensitivity enhancement and NOE measurements. J Am Chem Soc 115: 12593–12594, 1993Google Scholar
  31. 31.
    Fushman D, Cahill S, Cowburn D: The main-chain dynamics of the dynamin pleckstrin homology (PH) domain in solution: Analysis of 15N relaxation with monomer/dimer equilibration. J Mol Biol 266: 173–194, 1997Google Scholar
  32. 32.
    Lipari G, Szabo A: Model-free approach to the interpretation of nuclear magnetic resonance relaxation in macromolecules. 1. Theory and range of validity. J Am Chem Soc 104: 4546–4559, 1982Google Scholar
  33. 33.
    Lipari G, Szabo A: Model-free approach to the interpretation of nuclear magnetic resonance relaxation in macromolecules. 2. Analysis of experimental results. J Am Chem Soc 104: 4559–4570, 1982Google Scholar
  34. 34.
    Clore GM, Szabo A, Bax A, Kay LE, Driscoll PC, Wingfield PT, Gronenborn AM: Deviations from the simple two-parameter model-free approach to the interpretation of 15N nuclear magnetic relaxation of proteins. J Am Chem Soc 112: 4989–4991, 1990Google Scholar
  35. 35.
    Garcia de la Torre J, Bloomfield V: Hydrodynamic properties of complex rigid, biological macromolecules: Theory and applications. Quart Rev Biophys 14: 81–139, 1981Google Scholar
  36. 36.
    Tjandra N, Feler SE, Pastor RW, Bax A: Rotational diffusion anisotropy of human ubiquitin from 15N NMR relaxation. J Am Chem Soc 117: 12562–12566, 1995Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Christian Lücke
    • 1
  • David Fushman
    • 2
  • Christian Ludwig
    • 1
  • James A. Hamilton
    • 3
  • James C. Sacchettini
    • 4
  • Heinz Rüterjans
    • 1
  1. 1.Institut für Biophysikalische ChemieJ.W. Goethe-UniversitätFrankfurtGermany
  2. 2.The Rockefeller UniversityNew YorkUSA
  3. 3.Department of BiophysicsBoston University School of MedicineBostonUSA
  4. 4.Department of Biochemistry and BiophysicsTexas A&M UniversityCollege StationUSA

Personalised recommendations