Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin, China.
Lett Appl Microbiol. 2010 Oct;51(4):400-5. doi: 10.1111/j.1472-765X.2010.02911.x.
This work aims to investigate the binding capability of certain domains at N terminus of the protein NP_785232 of Lactobacillus plantarum to Caco-2 cells and to test the usage of affinity chromatography to isolate the human mucus component that interacts with them.
Recombinant proteins containing the first and both the first and second domains at N terminus of NP_785232 fused to a His tag were constructed and used to bind the Caco-2 cells. The interacting molecule from human mucus was isolated by affinity chromatography through immobilizing the recombinant proteins onto a Sepharose matrix. It was found both recombinant proteins could block the adhesion of Lact. plantarum to Caco-2 cells and bind to a human mucus component.
The first and both the first and second domains at N terminus of the protein NP_785232 have the capability to adhere Caco-2 cells and by affinity chromatography, an interacting UV-absorbing component from human mucus was isolated.
The protein domains characterized in this study may be displayed on probiotics to promote adhesion, and further characterization of the human mucus component might be helpful to identify host factors required for prolonging probiotics persistence in the gastrointestinal tract.
本研究旨在探究植物乳杆菌 NP_785232 蛋白 N 端前两个结构域与 Caco-2 细胞的结合能力，并通过亲和层析技术分离与这些结构域相互作用的人黏液成分。
构建了含有 NP_785232 蛋白 N 端前两个结构域和整个 N 端结构域并融合 His 标签的重组蛋白，并用于与 Caco-2 细胞结合。通过将重组蛋白固定在 Sepharose 基质上，通过亲和层析从人黏液中分离出与它们相互作用的分子。结果发现，两种重组蛋白均能阻止植物乳杆菌与 Caco-2 细胞的黏附，并与一种人黏液成分结合。
NP_785232 蛋白 N 端前两个结构域具有黏附 Caco-2 细胞的能力，通过亲和层析可分离出人黏液中与它们相互作用的紫外吸收成分。
本研究鉴定的蛋白结构域可能被展示在益生菌上以促进黏附，进一步鉴定人黏液成分有助于确定延长益生菌在胃肠道中持久性所需的宿主因素。
