title: Engineered xyloglucan specificity in a carbohydrate-binding module.
abstract: The field of plant cell wall biology is constantly growing and consequently so is the need for more sensitive and specific probes for individual wall components. Xyloglucan is a key polysaccharide widely distributed in the plant kingdom in both structural and storage tissues that exist in both fucosylated and non-fucosylated variants. Presently, the only xyloglucan marker available is the monoclonal antibody CCRC-M1 that is specific to terminal alpha-1,2-linked fucosyl residues on xyloglucan oligo- and polysaccharides. As a viable alternative to searches for natural binding proteins or creation of new monoclonal antibodies, an approach to select xyloglucan-specific binding proteins from a combinatorial library of the carbohydrate-binding module, CBM4-2, from xylanase Xyn10A of Rhodothermus marinus is described. Using phage display technology in combination with a chemoenzymatic method to anchor xyloglucan to solid supports, the selection of xyloglucan-binding modules with no detectable residual wild-type xylan and beta-glucan-binding ability was achieved.
authors.affiliation: Department of Immunotechnology, Lund University, BMC D13, SE-221 84 Lund, Sweden.
authors.forename: Lavinia Cicortas
authors.initials: LC
authors.lastname: Gunnarsson
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.forename: Cedric
authors.initials: C
authors.lastname: Montanier
authors.forename: Eva Nordberg
authors.initials: EN
authors.lastname: Karlsson
authors.forename: Harry
authors.initials: H
authors.lastname: Brumer
authors.forename: Mats
authors.initials: M
authors.lastname: Ohlin
mesh_terms: D000595:Amino Acid Sequence; D001710:Biotin / Q000737:chemistry; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D005643:Fucose / Q000737:chemistry; D005936:Glucans / Q000737:chemistry*; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D015202:Protein Engineering; D045245:Rhodothermus / Q000201:enzymology*; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity; D014990:Xylans / Q000737:chemistry*
chemical_list: D005936:Glucans; D019151:Peptide Library; D014990:Xylans; D005643:Fucose; C029353:xyloglucan; D001710:Biotin; D043364:Endo-1,4-beta Xylanases