MINORITY BIOMEDICAL RESEARCH PROGRAM--BENNETT COLLEGE

描述（改编自应用程序）：许多重要的细菌具有 除了典型的革兰氏阳性细胞外， 或阴性细胞壁。 这些可能不稳定的S层有助于 生物在恶劣的环境条件下生存。 条件 这些层的存在和功能并不清楚 明白 蛇形水螺菌具有这样的S层，因此 被用作研究这种细菌保护方法的模型。 的 PI证明了A. serpens由一个数组组成， 六角单元 化学上，这些单元由单一的酸性 糖蛋白（SG）。 合成的SG非共价组装成 细胞壁表面的S层。 S层的组装 提高了生物体的存活指数。 关于S层形成的许多机制方面仍然没有 知道的 PI目前正在研究聚糖部分的化学性质 的SG。 此外，对该化合物的物理化学性质知之甚少。 构象决定细胞壁大分子模板。 这个项目 试图扩展目前的研究，在分子水平上， (1)SG所对应的细胞包膜模板的确切性质 连接，和（2）聚糖部分和模板构象在 粘合-粘附过程。 确定模板将通过以下方式实现： 从细胞壁中分离大分子 模板，通过蛋白质印迹法将它们转移到合适的固体上 基质，并测试天然和去糖基化SG结合基质的能力。 在已知增强S层的条件下分离的大分子 阵 将使用兔SG特异性抗体评估结合 和碱性磷酸酶结合的抗兔抗血清。

Description (Adapted from Application): Many important bacteria possess surface layers (S-layers) that are additional to their typical Gram-positive or -negative cell wall. These S-layers, which may be unstable, aid the organisms in surviving adverse environmental conditions. The conditions under which these layers are present and functional are not clearly understood. Aquaspirillum serpens possesses such an S-layer and therefore is being used as a model to study this method of bacterial protection. The PI has shown that the S-layer of A. serpens is made up of an array of hexagonal units. Chemically, these units are composed of a single acidic glycoprotein (SG). The synthesized SG non-covalently assembles into the S-layer on the surface of the cell wall. The assembly of the S-layer increases the survival index of the organism. Many mechanistic aspects about the formation of the S-layer are still not known. The PI is currently investigating the chemistry of the glycan moiety of the SG. Also, little is known about the physicochemical nature of the conformation-determining cell wall macromolecular template. This project seeks to extend the current studies to investigate, at the molecular level, (1) the exact nature of the cell envelope template to which the SG is attached, and (2) the role of the glycan moiety and template conformation in the binding-adhesion process. Identifying the template will be achieved by electrophoretically separating the macromolecules from the cell wall template, transferring them by Western blotting onto a suitable solid matrix, and testing the ability of native and deglycosylated SG to bind to the separated macromolecules under conditions known to enhance S-layer formation. The binding will be assessed using rabbit SG-specific antibodies and alkaline phosphatase-conjugated antirabbit antiserum.