title: Structural motif screening reveals a novel, conserved carbohydrate-binding surface in the pathogenesis-related protein PR-5d.
abstract: BACKGROUND
  Aromatic amino acids play a critical role in protein-glycan interactions. Clusters of surface aromatic residues and their features may therefore be useful in distinguishing glycan-binding sites as well as predicting novel glycan-binding proteins. In this work, a structural bioinformatics approach was used to screen the Protein Data Bank (PDB) for coplanar aromatic motifs similar to those found in known glycan-binding proteins.
  
  
  RESULTS
  The proteins identified in the screen were significantly associated with carbohydrate-related functions according to gene ontology (GO) enrichment analysis, and predicted motifs were found frequently within novel folds and glycan-binding sites not included in the training set. In addition to numerous binding sites predicted in structural genomics proteins of unknown function, one novel prediction was a surface motif (W34/W36/W192) in the tobacco pathogenesis-related protein, PR-5d. Phylogenetic analysis revealed that the surface motif is exclusive to a subfamily of PR-5 proteins from the Solanaceae family of plants, and is absent completely in more distant homologs. To confirm PR-5d's insoluble-polysaccharide binding activity, a cellulose-pulldown assay of tobacco proteins was performed and PR-5d was identified in the cellulose-binding fraction by mass spectrometry.
  
  
  CONCLUSIONS
  Based on the combined results, we propose that the putative binding site in PR-5d may be an evolutionary adaptation of Solanaceae plants including potato, tomato, and tobacco, towards defense against cellulose-containing pathogens such as species of the deadly oomycete genus, Phytophthora. More generally, the results demonstrate that coplanar aromatic clusters on protein surfaces are a structural signature of glycan-binding proteins, and can be used to computationally predict novel glycan-binding proteins from 3 D structure.
authors.affiliation: Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Doxey
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Cheng
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Moffatt
authors.forename: Brendan J
authors.initials: BJ
authors.lastname: McConkey
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D024322:Amino Acids, Aromatic; D050260:Carbohydrate Metabolism; D002482:Cellulose / Q000737:chemistry / Q000378:metabolism; D019295:Computational Biology / Q000379:methods*; D017124:Conserved Sequence; D030562:Databases, Protein; D016002:Discriminant Analysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D012995:Solubility; D013045:Species Specificity; D014026:Nicotiana / Q000145:classification
chemical_list: D024322:Amino Acids, Aromatic; D010940:Plant Proteins; C053376:pathogenesis-related proteins, plant; D002482:Cellulose