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半乳甘露聚糖薄膜作为固定刀豆球蛋白 A 和/或登革热病毒的载体。

Galactomannan thin films as supports for the immobilization of Concanavalin A and/or dengue viruses.

机构信息

Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba, PR, Brazil.

出版信息

Int J Biol Macromol. 2012 Jan 1;50(1):88-94. doi: 10.1016/j.ijbiomac.2011.10.005. Epub 2011 Oct 15.

DOI:10.1016/j.ijbiomac.2011.10.005
PMID:22020153
Abstract

The immobilization of the glucose/mannose-binding lectin from Concanavalia ensiformis seeds (ConA) onto a monolayer made of a galactomannan extracted from Leucaena leucocephala seeds (GML), which was adsorbed onto - amino-terminated surfaces, was investigated by means of ellipsometry and atomic force microscopy. The mean thickness of GML monolayer, which polysaccharide consists of linear 1→4-linked β-D-mannopyranosil units partially substituted at C-6 by α-D-galactopyranosyl units, amounted to (1.5±0.2) nm. ConA molecules adsorbed onto GML surfaces forming (2.0±0.5) nm thick layers. However, in the presence of mannose the adsorption failed, indicating that ConA binding sites were blocked by mannose and were no longer available for mannose units present in the GML backbone. The GML film was also used as support for the adsorption of three serotypes of dengue virus particles (DENV-1, DENV-2 and DENV-3), where DENV-2 formed the thickest film (4±2) nm. The adsorbed layer of DENV-2 onto ConA-covered GML surfaces presented mean thickness values similar to that determined for DENV-2 onto bare GML surfaces. The addition of free mannose units prevented DENV-2 adsorption onto ConA-covered GML films by ~50%, suggesting competition between virus and mannose for ConA binding sites. This finding suggests that if ConA is also adsorbed to GML surface and its binding site is blocked by free mannose, virus particles are able to recognized GML mannose unities substituted by galactose. Interactions between polysaccharides thin films, proteins, and viruses are of great relevance since they can provide basis for the development of biotechnological devices. These results indicate that GML is a potential polysaccharide for biomaterials development, as those could involve interactions between ConA in immune system and viruses.

摘要

用椭圆偏振法和原子力显微镜研究了 Concanavalia ensiformis 种子(ConA)葡萄糖/甘露糖结合凝集素固定在半乳糖甘露聚糖(GML)单层上的情况,GML 是从 Leucaena leucocephala 种子中提取的,并吸附在 - 氨基末端表面上。多糖由线性 1→4 连接的β-D-甘露吡喃糖基单元组成,部分在 C-6 上被α-D-半乳糖吡喃糖基单元取代,GML 单层的平均厚度为(1.5±0.2)nm。ConA 分子吸附在 GML 表面上,形成(2.0±0.5)nm 厚的层。然而,在甘露糖存在的情况下,吸附失败,表明 ConA 结合位点被甘露糖封闭,不再可用于 GML 主链中的甘露糖单元。GML 膜也被用作三种登革热病毒颗粒(DENV-1、DENV-2 和 DENV-3)的吸附载体,其中 DENV-2 形成最厚的膜(4±2)nm。DENV-2 吸附到 ConA 覆盖的 GML 表面上的吸附层的平均厚度值与 DENV-2 吸附到裸露 GML 表面上的平均厚度值相似。游离甘露糖单元的加入阻止了 DENV-2 吸附到 ConA 覆盖的 GML 膜上约 50%,表明病毒和甘露糖与 ConA 结合位点之间存在竞争。这一发现表明,如果 ConA 也被吸附到 GML 表面,并且其结合位点被游离甘露糖封闭,那么病毒颗粒能够识别被半乳糖取代的 GML 甘露糖单元。多糖薄膜、蛋白质和病毒之间的相互作用非常重要,因为它们可以为生物技术设备的发展提供基础。这些结果表明,GML 是一种有潜力的生物材料开发多糖,因为它可以涉及免疫系统中的 ConA 与病毒之间的相互作用。

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