title: Substituent-specific antibody against glucuronoxylan reveals close association of glucuronic acid and acetyl substituents and distinct labeling patterns in tree species.
abstract: Immunolabeling can be used to locate plant cell wall carbohydrates or other components to specific cell types or to specific regions of the wall. Some antibodies against xylans exist; however, many partly react with the xylan backbone and thus provide limited information on the type of substituents present in various xylans. We have produced a monoclonal antibody which specifically recognizes glucopyranosyl uronic acid (GlcA), or its 4-O-methyl ether (meGlcA), substituents in xylan and has no cross-reactivity with linear or arabinofuranosyl-substituted xylans. The UX1 antibody binds most strongly to (me)GlcA substitutions at the non-reducing ends of xylan chains, but has a low cross-reactivity with internal substitutions as well, at least on oligosaccharides. The antibody labeled plant cell walls from both mono- and dicotyledons, but in most tissues an alkaline pretreatment was needed for antibody binding. The treatment removed acetyl groups from xylan, indicating that the vicinity of glucuronic acid substituents is also acetylated. The novel labeling patterns observed in the xylem of tree species suggested that differences within the cell wall exist both in acetylation degree and in glucuronic acid content.
authors.affiliation: Department of Food and Environmental Sciences, University of Helsinki, PO Box 27, 00014, Helsinki, Finland. sanna.koutaniemi@helsinki.fi
authors.forename: Sanna
authors.initials: S
authors.lastname: Koutaniemi
authors.forename: Fabienne
authors.initials: F
authors.lastname: Guillon
authors.forename: Olivier
authors.initials: O
authors.lastname: Tranquet
authors.forename: Brigitte
authors.initials: B
authors.lastname: Bouchet
authors.forename: Päivi
authors.initials: P
authors.lastname: Tuomainen
authors.forename: Liisa
authors.initials: L
authors.lastname: Virkki
authors.forename: Henriette L
authors.initials: HL
authors.lastname: Petersen
authors.forename: William G T
authors.initials: WG
authors.lastname: Willats
authors.forename: Luc
authors.initials: L
authors.lastname: Saulnier
authors.forename: Maija
authors.initials: M
authors.lastname: Tenkanen
mesh_terms: D000107:Acetylation; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000918:Antibody Specificity; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005965:Glucuronates / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006825:Hybridomas; D007114:Immunization; D009682:Magnetic Resonance Spectroscopy; D019684:Magnoliopsida / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008856:Microscopy, Fluorescence; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results; D014990:Xylans / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D052584:Xylem / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000911:Antibodies, Monoclonal; D005965:Glucuronates; D009844:Oligosaccharides; D014990:Xylans; C570991:xylooligosaccharide; C036035:4-O-methylglucuronic acid; C038910:glucuronoxylan