title: Solution structure of Eucommia antifungal peptide: a novel structural model distinct with a five-disulfide motif.
abstract: The three-dimensional structure in aqueous solution of Eucommia antifungal peptide 2 (EAFP2) from Eucommia ulmoides Oliv was determined using (1)H NMR spectroscopy. EAFP2 is a newly discovered 41-residue peptide distinct with a five-disulfide cross-linked motif. This peptide exhibits chitin-binding activity and inhibitory effects on the growth of cell wall chitin-containing fungi and chitin-free fungi. The structure was calculated by using torsion angle dynamic simulated annealing with a total of 614 distance restraints and 16 dihedral restraints derived from NOESY and DQF-COSY spectra, respectively. The five disulfide bonds were assigned from preliminary structures using a statistical analysis of intercystinyl distances. The solution structure of EAFP2 is presented as an ensemble of 20 conformers with a backbone RMS deviation of 0.65 (+/-0.13) A for the well-defined Cys3-Cys39 segment. The tertiary structure of EAFP2 represents the first five-disulfide cross-linked structural model of the plant antifungal peptide. EAFP2 adopts a compact global fold composed of a 3(10) helix (Cys3-Arg6), an alpha-helix (Gly26-Cys30), and a three-strand antiparallel beta-sheet (Cys16-Ser18, Tyr22-Gly24, and Arg36-Cys37). The tertiary structure of EAFP2 shows a chitin-binding domain (residues 11-30) with a hydrophobic face and a characteristic sector formed by the N-terminal 10 residues and the C-terminal segment cross-linked through the unique disulfide bond Cys7-Cys37, which brings all four positively charged residues (Arg6, Arg9, Arg36, and Arg40) onto a cationic face. On the basis of such a structural feature, the possible structural basis for the functional properties of EAFP2 is discussed.
authors.affiliation: Center for Structural and Molecular Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, PR China.
authors.forename: Ren-Huai
authors.initials: RH
authors.lastname: Huang
authors.forename: Ye
authors.initials: Y
authors.lastname: Xiang
authors.forename: Guan-Zhong
authors.initials: GZ
authors.lastname: Tu
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Da-Cheng
authors.initials: DC
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides; D001665:Binding Sites; D004220:Disulfides / Q000737:chemistry*; D031283:Eucommiaceae / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D037121:Plant Lectins / Q000737:chemistry / Q000235:genetics; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D004220:Disulfides; C488625:EAFP2 peptide, Eucommia ulmoides; D010455:Peptides; D037121:Plant Lectins; D010940:Plant Proteins; C045875:hevein