The mechanism of crosslinking by proteins that create adhesive gels

描述（由申请人提供）：各种各样的动物使用凝胶为基础的胶水来创建强附着物。这些凝胶由大约97%的水组成，但它们可以产生附着物，其强度接近其他海洋动物的固体粘合剂。在潮湿环境中用高度可变形的水基胶粘剂形成如此牢固的附着物的能力将对医用胶粘剂非常有用。因此，确定这些粘合剂的功能是很重要的。软体动物的胶基胶的关键成分是胶蛋白的存在，它非特异性地使胶变硬。拟议研究的目标是确定胶水蛋白的机制。据推测，胶蛋白通过离子和疏水相互作用的组合交联较大的聚合物。将使用三种方法来测试这一点。首先，从陆生蜗牛或蛞蝓中提取的胶蛋白将被纯化，然后添加到商业凝胶形成聚合物中。将在各种干扰特定相互作用的条件下测量胶蛋白的凝胶硬化能力。其次，柱层析法将用于测量胶蛋白在相同条件下与不同底物相互作用的能力。最后，将使用质谱法和聚合酶链反应（PCR）方法确定其中一个主要粘合蛋白的一级序列。这将有助于确定可能的交联区域，并进一步了解它们的功能。这项研究的长期目标是提供足够的信息，以设计一种实用的粘合剂使用仿生方法。这与公共卫生有关，因为显然需要改进医用粘合剂。一旦了解了动物使用的非凡的天然粘合剂，就有可能设计出具有类似性质的粘合剂，可用于各种医疗应用。

DESCRIPTION (provided by applicant): A wide variety of animals use gel-based glues to create strong attachments. These gels consist of roughly 97% water, yet they can create attachments that approach the strength of the solid cements of other marine animals. The ability to form such strong attachments in wet environments with a highly deformable, water- based adhesive would be highly useful for a medical adhesive. Thus, it is important to determine how these adhesives function. A key component of the gel-based glues of molluscs is the presence of glue proteins, which non-specifically stiffen gels. The goal of the proposed research is to determine the mechanism by which the glue proteins do this. It is hypothesized that the glue proteins crosslink larger polymers through a combination of ionic and hydrophobic interactions. Three approaches will be used to test this. First, the glue proteins from terrestrial snails or slugs will be purified, then added to commercial gel-forming polymers. The gel-stiffening ability of the glue proteins will be measured under a variety of conditions that interfere with specific interactions. Second, column chromatography will be used to measure the ability of the glue proteins to interact with different substrates under the same kind of conditions. Finally, mass spectrometry and Polymerase Chain Reaction (PCR) based methods will be used to determine the primary sequence of one of the main glue proteins. This will help identify possible cross-linking regions and give further insight into how they may function. The long-term goal of this research is to provide sufficient information to design a practical adhesive using a biomimetic approach. This is relevant to public health because there is a clear need for improved medical adhesives. Once the remarkable natural adhesives used by animals are understood, it may be possible to design adhesives with similar properties that could be used in a wide variety of medical applications.