Comparison of the three-dimensional structures of grape TLP.

<p>A) Mapping of the electrostatic potentials on the molecular surface of the grape TLP F2/4JRU. The negative potentials and positive potentials are colored red and blue, respectively. Neutral surfaces are white. Arrow indicates the acidic cleft region located in between domains I and II. B) Superposition of the backbone representations of chain A (in red) and B (blue) of I/4L5H. TM-align data showed that the two chains of I/4L5H have RMSD of 0.14 Å, and a TM-score of 0.99925. C) Superposition of the backbone representations of chain A (in red) and B (blue) of H2/4MBT. TM-align data showed that the two chains of H2/4MBT have RMSD of 0.13 Å, and a TM-score of 0.99939. D) Superposition of the secondary structure of I/4L5H chain A (in purple) and F2/4JRU (in blue). Arrows indicate the two loop regions showing the largest differences between the two structures. RMSD between 193 atom pairs is 0.505 Å (calculated with UCSF Chimera MatchMaker function). E) Detail of differences in loop regions in Domain I. F) Detail of differences in loop region in Domain II. Cysteine are shown in yellow, and disulfide bonds are formed between residues 140–213 and 183–178 in F2/4JRU, and residues 136–210 and 180–173 in I/4L5H, as indicated by the asterisks.</p

Three-dimensional structure of protein F2/4JRU.

<p>A) Ribbon diagram of the overall three-dimensional structure of protein F2/4JRU. The protein consists of three structural domains: a central core domain I built from a β-sandwich of two sheets of six (front) and five (back) β-strands, flanked by two shorted domains II (where the 5 α-helixes and β-strand 11 and 12 are located) and III (where two β-strands - 5 and 6 - and a turn form two looping regions). The eight disulphide bridges are shown in yellow. B) Superposition of the backbone representations of the grape TLP F2/4JRU (in orange) and banana TLP 1Z3Q (in blue). TM-align data showed that the two proteins have RMSD of 0.78 Å, and a TM-score of 0.98 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113757#pone.0113757-Zhang1" target="_blank">[41]</a>. Significant differences in loops' structures are indicated by arrows, and numbers denote residues in F2/4JRU. C) and D) Mapping of the surface hydrophobicity of the grape TLP F2/4JRU, front view (same as Figure 2A) and back view (rotated approximately 180°) respectively. Hydrophobicity continuum from orange to blue representing hydrophobic to hydrophilic. E) Topology of protein F2/4JRU showing the residues included in each β-strand or α-helix.</p

SDS-PAGE of purified proteins under reducing and non-reducing conditions.

<p>Proteins (∼2 µg per lane) were reduced with 5% BME or loaded on a 12% gel in non-reducing conditions and subjected to SDS-PAGE.</p

Data collection and processing.

<p>Values for the highest resolution shell are shown in parentheses.</p><p>Data collection and processing.</p