Skip to main content
PMC home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1996 Jan;5(1):13–23. doi: 10.1002/pro.5560050103

Structure in solution of a four-helix lipid binding protein.

B Heinemann 1, K V Andersen 1, P R Nielsen 1, L M Bech 1, F M Poulsen 1
PMCID: PMC2143251  PMID: 8771192

Abstract

Because of the low solubility of lipids in water, intercellular and intracellular pathways of lipid transfer are necessary, e.g., for membrane formation. The mechanism by which lipids in vivo are transported from their site of biogenesis (endoplasmatic reticulum and the chloroplasts) to their place of action is unknown. Several small plant proteins with the ability to mediate transfer of radiolabeled phospholipids in vitro from liposomal donor membranes to mitochondrial and chloroplast acceptor membranes have been isolated, and a protein with this ability, the nonspecific lipid transfer protein (nsLTP) isolated from barley seeds (bLTP), has been studied here. The structure and the protein lipid interactions of lipid transfer proteins are relevant for the understanding of their function, and here we present the three-dimensional structure in solution of bLTP as determined by NMR spectroscopy. The 1H NMR spectrum of the 91-residue protein was assigned for more than 97% of the protein 1H atoms, and the structure was calculated on the basis of 813 distance restraints from 1H-1H nuclear Overhauser effects, four disulfide bond restraints, from dihedral angle restraints for 66 phi-angles, 61 chi 1 angles, and 2 chi 2 angles, and from 31 sets of hydrogen bond restraints. The solution structure of bLTP consists of four well-defined alpha-helices A-D (A, Cys 3-Gly 19; B, Gly 25-Ala 38; C, Arg 44-Gly 57; D, Leu 63-Cys 73), separated by three short loops that are less well defined and concluded by a well defined C-terminal peptide segment with no observable regular secondary structure. For the 17 structures that are used to represent the solution structure of bLTP, the RMS deviation to an average structure is 0.63 A +/- 0.04 A for backbone atoms and 0.93 A +/- 0.06 A for all heavy atoms. The secondary structure elements and their locations in the sequence resemble those of nsLTP from two other plant species, wheat and maize, whose structures were previously determined (Gincel E et al, 1995, Eur J Biochem 226:413-422; Shin DH et al, 1995, Structure 3:189-199). In bLTP, the residues analogous to those in maize nsLTP that constitute the palmitate binding site are forming a similar hydrophobic cavity and a potential acyl group binding site. Analysis of the solution structure of bLTP and bLTP in complex with a ligand might provide information on the conformational changes in the protein upon ligand binding and subsequently provide information on the mode of ligand uptake and release. In this work, we hope to establish a foundation for further work of determining the solution structure of bLTP in complex with palmitoyl coenzyme A, which is a suitable ligand, and subsequently to outline the mode of ligand binding.

Full Text

The Full Text of this article is available as a PDF (6.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersen K. V., Poulsen F. M. The three-dimensional structure of acyl-coenzyme A binding protein from bovine liver: structural refinement using heteronuclear multidimensional NMR spectroscopy. J Biomol NMR. 1993 May;3(3):271–284. doi: 10.1007/BF00212514. [DOI] [PubMed] [Google Scholar]
  2. Andersen K. V., Poulsen F. M. Three-dimensional structure in solution of acyl-coenzyme A binding protein from bovine liver. J Mol Biol. 1992 Aug 20;226(4):1131–1141. doi: 10.1016/0022-2836(92)91057-v. [DOI] [PubMed] [Google Scholar]
  3. Bernhard W. R., Somerville C. R. Coidentity of putative amylase inhibitors from barley and finger millet with phospholipid transfer proteins inferred from amino acid sequence homology. Arch Biochem Biophys. 1989 Mar;269(2):695–697. doi: 10.1016/0003-9861(89)90154-9. [DOI] [PubMed] [Google Scholar]
  4. Désormeaux A., Blochet J. E., Pézolet M., Marion D. Amino acid sequence of a non-specific wheat phospholipid transfer protein and its conformation as revealed by infrared and Raman spectroscopy. Role of disulfide bridges and phospholipids in the stabilization of the alpha-helix structure. Biochim Biophys Acta. 1992 May 22;1121(1-2):137–152. doi: 10.1016/0167-4838(92)90347-g. [DOI] [PubMed] [Google Scholar]
  5. Gincel E., Simorre J. P., Caille A., Marion D., Ptak M., Vovelle F. Three-dimensional structure in solution of a wheat lipid-transfer protein from multidimensional 1H-NMR data. A new folding for lipid carriers. Eur J Biochem. 1994 Dec 1;226(2):413–422. doi: 10.1111/j.1432-1033.1994.tb20066.x. [DOI] [PubMed] [Google Scholar]
  6. Kabsch W., Sander C. Dictionary of protein secondary structure: pattern recognition of hydrogen-bonded and geometrical features. Biopolymers. 1983 Dec;22(12):2577–2637. doi: 10.1002/bip.360221211. [DOI] [PubMed] [Google Scholar]
  7. Kader J. C., Julienne M., Vergnolle C. Purification and characterization of a spinach-leaf protein capable of transferring phospholipids from liposomes to mitochondria or chloroplasts. Eur J Biochem. 1984 Mar 1;139(2):411–416. doi: 10.1111/j.1432-1033.1984.tb08020.x. [DOI] [PubMed] [Google Scholar]
  8. Kjaer M., Andersen K. V., Poulsen F. M. Automated and semiautomated analysis of homo- and heteronuclear multidimensional nuclear magnetic resonance spectra of proteins: the program Pronto. Methods Enzymol. 1994;239:288–307. doi: 10.1016/s0076-6879(94)39010-x. [DOI] [PubMed] [Google Scholar]
  9. Kragelund B. B., Andersen K. V., Madsen J. C., Knudsen J., Poulsen F. M. Three-dimensional structure of the complex between acyl-coenzyme A binding protein and palmitoyl-coenzyme A. J Mol Biol. 1993 Apr 20;230(4):1260–1277. doi: 10.1006/jmbi.1993.1240. [DOI] [PubMed] [Google Scholar]
  10. Ludvigsen S., Andersen K. V., Poulsen F. M. Accurate measurements of coupling constants from two-dimensional nuclear magnetic resonance spectra of proteins and determination of phi-angles. J Mol Biol. 1991 Feb 20;217(4):731–736. doi: 10.1016/0022-2836(91)90529-f. [DOI] [PubMed] [Google Scholar]
  11. Molina A., García-Olmedo F. Developmental and pathogen-induced expression of three barley genes encoding lipid transfer proteins. Plant J. 1993 Dec;4(6):983–991. doi: 10.1046/j.1365-313x.1993.04060983.x. [DOI] [PubMed] [Google Scholar]
  12. Molina A., Segura A., García-Olmedo F. Lipid transfer proteins (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens. FEBS Lett. 1993 Jan 25;316(2):119–122. doi: 10.1016/0014-5793(93)81198-9. [DOI] [PubMed] [Google Scholar]
  13. Newcomer M. E., Jones T. A., Aqvist J., Sundelin J., Eriksson U., Rask L., Peterson P. A. The three-dimensional structure of retinol-binding protein. EMBO J. 1984 Jul;3(7):1451–1454. doi: 10.1002/j.1460-2075.1984.tb01995.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nilges M., Clore G. M., Gronenborn A. M. Determination of three-dimensional structures of proteins from interproton distance data by hybrid distance geometry-dynamical simulated annealing calculations. FEBS Lett. 1988 Mar 14;229(2):317–324. doi: 10.1016/0014-5793(88)81148-7. [DOI] [PubMed] [Google Scholar]
  15. Nilsson L., Clore G. M., Gronenborn A. M., Brünger A. T., Karplus M. Structure refinement of oligonucleotides by molecular dynamics with nuclear Overhauser effect interproton distance restraints: application to 5' d(C-G-T-A-C-G)2. J Mol Biol. 1986 Apr 5;188(3):455–475. doi: 10.1016/0022-2836(86)90168-3. [DOI] [PubMed] [Google Scholar]
  16. Plant G. W., Harvey A. R., Chirila T. V. Axonal growth within poly (2-hydroxyethyl methacrylate) sponges infiltrated with Schwann cells and implanted into the lesioned rat optic tract. Brain Res. 1995 Feb 6;671(1):119–130. doi: 10.1016/0006-8993(94)01312-6. [DOI] [PubMed] [Google Scholar]
  17. Takishima K., Watanabe S., Yamada M., Suga T., Mamiya G. Amino acid sequences of two nonspecific lipid-transfer proteins from germinated castor bean. Eur J Biochem. 1988 Nov 1;177(2):241–249. doi: 10.1111/j.1432-1033.1988.tb14368.x. [DOI] [PubMed] [Google Scholar]
  18. Tanaka T., Ames J. B., Harvey T. S., Stryer L., Ikura M. Sequestration of the membrane-targeting myristoyl group of recoverin in the calcium-free state. Nature. 1995 Aug 3;376(6539):444–447. doi: 10.1038/376444a0. [DOI] [PubMed] [Google Scholar]
  19. Tchang F., This P., Stiefel V., Arondel V., Morch M. D., Pages M., Puigdomenech P., Grellet F., Delseny M., Bouillon P. Phospholipid transfer protein: full-length cDNA and amino acid sequence in maize. Amino acid sequence homologies between plant phospholipid transfer proteins. J Biol Chem. 1988 Nov 15;263(32):16849–16855. [PubMed] [Google Scholar]
  20. Terras F. R., Goderis I. J., Van Leuven F., Vanderleyden J., Cammue B. P., Broekaert W. F. In Vitro Antifungal Activity of a Radish (Raphanus sativus L.) Seed Protein Homologous to Nonspecific Lipid Transfer Proteins. Plant Physiol. 1992 Oct;100(2):1055–1058. doi: 10.1104/pp.100.2.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tsuboi S., Osafune T., Tsugeki R., Nishimura M., Yamada M. Nonspecific lipid transfer protein in castor bean cotyledon cells: subcellular localization and a possible role in lipid metabolism. J Biochem. 1992 Apr;111(4):500–508. doi: 10.1093/oxfordjournals.jbchem.a123787. [DOI] [PubMed] [Google Scholar]
  22. Vergnolle C., Arondel V., Tchang F., Grosbois M., Guerbette F., Jolliot A., Kader J. C. Synthesis of phospholipid transfer proteins from maize seedlings. Biochem Biophys Res Commun. 1988 Nov 30;157(1):37–41. doi: 10.1016/s0006-291x(88)80007-x. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES