MINORITY BIOMEDICAL RESEARCH PROGRAM--BENNETT COLLEGE

描述(改编自应用)：许多重要的细菌都具有 典型革兰氏阳性菌以外的表层(S层) 或-阴性细胞壁。这些可能不稳定的S层，有助于 在不利的环境条件下生存的有机体。这些条件 这些层的存在和功能并不清楚 明白了。蛇水藻具有这样一层S层，因此 正被用作研究这种细菌保护方法的模型。这个 Pi已经证明，蛇的S层是由一组 六角形单位。在化学上，这些单元是由一个单一的酸性 糖蛋白(SG)。合成的SG以非共价方式组装成 S--细胞壁表面的一层。S层的集结 增加有机体的生存指数。 关于S层形成的许多机制方面仍然不是 为人所知。PI目前正在研究葡聚糖部分的化学 总司令的。此外，人们对这种物质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，在分子水平上调查， (1)SG所属的单元格包络模板的确切性质 以及(2)糖链部分和模板构象在 结合-粘合过程。识别模板将通过以下方式实现 电泳法将大分子从细胞壁中分离出来 模板，通过Western blotting将它们转移到合适的固体上 并检测天然的和脱糖的SG结合的能力 分离的大分子在已知条件下对S层有增强作用 队形。结合将使用兔SG特异性抗体进行评估 碱性磷酸酶标记兔抗血清。

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.