Gunnarsson Lavinia Cicortas, Zhou Qi, Montanier Cedric, Karlsson Eva Nordberg, Brumer Harry, Ohlin Mats
Department of Immunotechnology, Lund University, BMC D13, SE-221 84 Lund, Sweden.
Glycobiology. 2006 Dec;16(12):1171-80. doi: 10.1093/glycob/cwl038. Epub 2006 Aug 10.
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.
植物细胞壁生物学领域在不断发展，因此对用于单个细胞壁成分的更灵敏、更特异的探针的需求也在增加。木葡聚糖是一种关键的多糖，广泛分布于植物界的结构组织和贮藏组织中，存在岩藻糖基化和非岩藻糖基化变体。目前，唯一可用的木葡聚糖标记物是单克隆抗体CCRC-M1，它对木葡聚糖寡糖和多糖上的末端α-1,2-连接的岩藻糖基残基具有特异性。作为寻找天然结合蛋白或制备新单克隆抗体的可行替代方法，本文描述了一种从海栖热袍菌木聚糖酶Xyn10A的碳水化合物结合模块CBM4-2的组合文库中筛选木葡聚糖特异性结合蛋白的方法。利用噬菌体展示技术结合化学酶法将木葡聚糖锚定到固体支持物上，实现了对无野生型木聚糖和β-葡聚糖结合能力残留的木葡聚糖结合模块的筛选。
