Insight into freeze resistance and proteostasis from protective proteins and small organic molecules of marine species

蛋白质在压力条件下可以解绕（展开）。因此，本研究旨在了解寒冷海洋和潮间带环境中的物种如何维持折叠蛋白。研究的重点是与冰结合并抑制其生长的抗冻蛋白，以及促进蛋白质适当折叠的小分子化学伴侣蛋白。对蓝贻贝抗冻蛋白进行纯化和表征。该蛋白和来自冬季比目鱼的两种抗冻蛋白将作为化学伴侣蛋白增强蛋白质折叠研究的模型。抗冻蛋白将是这项工作的理想模型，因为它们似乎具有不同的折叠限制。我们感兴趣的化学伴侣包括那些在贻贝中自然存在的化学伴侣和一些其他化合物。伴侣活性将使用蛋白质折叠的标准测试进行评估。在适当的情况下，将对蛋白质进行更详细的结构研究。这项工作将为贻贝提供一种新的具有特征的抗冻蛋白，并深入了解该物种的伴侣分子。本研究将确定冬季短比目鱼抗冻蛋白中引起淀粉样蛋白形成的参数，并指出化学伴侣预防淀粉样蛋白的可能性。这项工作还将显示冬季比目鱼的较长，高活性的抗冻蛋白在变性后是否形成淀粉样蛋白或非淀粉样蛋白聚集体，并使我们能够确定是否可以通过监护人来保持其折叠。总的来说，这项研究将为淀粉样蛋白的形成和化学伴侣的功能提供更好的理解。我们预期以下具体结果：(1)增强对极端海洋和潮间带环境中物种的分子水平理解；(2)这些物种使用的生物策略，可以人工应用于其他环境中的组织、细胞或蛋白质的稳定；(3)可用于预防或治疗蛋白质折叠疾病的化合物和过程（例如人类的几种神经系统疾病）。

Proteins can become unwound (unfolded) under stressful conditions. Therefore, this proposal aims to understand how species in cold ocean and intertidal environments maintain folded proteins. The focus is on antifreeze proteins, which bind to ice and inhibit its growth, and on chemical chaperones, which are small molecules that promote proper protein folding. The blue mussel antifreeze protein will be purified and characterized. This protein and two antifreeze proteins from winter flounder will then serve as models for the study of protein folding enhancement by chemical chaperones. The antifreeze proteins will be ideal models in this work because they appear to have divergent folding limitations. The chemical chaperones of interest include those that occur naturally in the mussel and a selection of other compounds. Chaperone activity will be evaluated using standard tests for protein folding. Where appropriate, there will be more detailed structural studies on the proteins. This work will provide a novel characterized antifreeze protein from the mussel and insight into the chaperoning molecules in this species. This work will define parameters causing amyloid formation in the short winter flounder antifreeze protein and indicate the possibility of amyloid prevention by chemical chaperones. This work will also show whether the longer, highly active antifreeze protein of winter flounder forms amyloid or non-amyloid aggregates upon denaturation and it will allow us to determine whether the long antifreeze protein can be chaperoned to maintain its fold. Overall, this research will provide greater understanding of amyloid formation and chemical chaperone function. We anticipate the following specific outcomes: (1) enhanced molecular-level understanding of the species that inhabit extreme marine and intertidal environments, (2) biological strategies used by these species that could be applied artificially for the stabilization of tissues, cells or proteins in other contexts, and (3) compounds and processes that could be used to prevent or treat diseases of protein folding (such as several neurological diseases in humans).