Protein Chaperone Nanoparticles - Abiotic Mimics of Heat Shock Proteins

蛋白质伴侣纳米颗粒 - 热休克蛋白的非生物模拟物

• 批准号: 1308363
• 负责人: Kenneth Shea
• 金额: $ 44万
• 依托单位: University of California-Irvine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1308363&HistoricalAwards=false
• 关键词: Protein; Chaperone; Nanoparticles; Abiotic; Mimics

This award by the Biomaterials program in the Division of Materials Research is to develop novel, low cost, abiotic, thermally responsive polyacrylamide hydrogel nanoparticles with chaperone-like activity to stabilize proteins at elevated temperatures. The chaperone-like function arises from two 'engineered' properties of the synthetic polymer nanoparticles: 1) an intrinsic antibody-like protein affinity; and 2) a lower critical solution temperature (LCST), which can be used to trap protein at elevated temperatures, preventing its aggregation and denaturation, and release of the protein upon cooling below the LCST without the need for additives and preserving the overall function of the protein in solution. The concept of creating 'smart' synthetic polymer molecular chaperones, where the antibody-like protein affinity of synthetic polymer nanoparticles is utilized to stabilize a target protein by inhibiting denaturation and/or protein-protein aggregation and the resulting loss of activity. The second 'smart' function of the synthetic polymer nanoparticles arises from its thermal response to temperature. The polymer is in a collapsed state at a temperature above its lower critical solution temperature, where it has protein binding affinity. These two unique polymer properties allow engineering the protein binding affinity of the nanoparticle so that it is switched on and off in response to temperature. One application would be a nanoparticle that sequesters and stabilizes a target protein at elevated temperatures, where the protein is vulnerable, but that has little or no affinity for the protein at room temperature so as to not impede the proteins function. A 'smart' material with this property would find applications by protecting proteins from denaturation. It could also help reduce the cost of protein therapeutics and diagnostics, which arise in part due to the retention of efficacy of the drugs during transportation and storage. This proposal will engage graduate students, undergraduates and high school students in an interdisciplinary research program that spans polymer synthesis, materials chemistry, biology and biotechnology. Additionally, the project will provide students with an opportunity to work with industrial collaborators on practical problems, which could have profound impact on the world-wide delivery of health care. Proteins are large biopolymers produced by all living systems. They play an essential role in all biological processes including regulatory processes, the immune system, catalysis of all biochemical reactions among other functions and properties. Despite their complexity, advances in molecular biology and biotechnology have made proteins readily available. These 'engineered' proteins are now produced and used on a large scale for applications that go far beyond their intended biological functions. These applications include therapeutics and diagnostics, and synthesis of value-added chemicals. Proteins, however, have several limitations. They are not stable when taken beyond physiological conditions of temperature and pH. In particular, they are often unstable at conditions above room temperature, and this temperature sensitivity can limit the use of proteins particularly where refrigeration facilities are not easily available. Maintaining protein functions depend on the availability of a network of cold storage facilities, which will provide optimal cold temperatures during transport, storage, and handling. Nature has evolved strategies to protect proteins from elevated temperatures. A class of biomacromolecules collectively referred to as heat shock proteins, 'protect' vulnerable proteins against loss of function at elevated temperatures. This proposed study is in developing simple, low cost synthetic polymers that can mimic the behavior of heat shock proteins. These 'engineered' synthetic polymer nanoparticles are being designed to 'protect' proteins from denaturation at elevated temperature by sequestering them. This proposal will provide educational and training opportunities to students from graduate to high school levels in an interdisciplinary research program that spans many scientific fields such as polymer synthesis, materials chemistry, biotechnology and biology. In addition, this project will provide students with opportunity to work with industrial collaborators and receive training not only in preparing them as the future generation of scientists but also providing them with opportunities to work in industry.

该奖项由材料研究部生物材料计划授予，旨在开发新型，低成本，非生物，热响应聚丙烯酰胺水凝胶纳米颗粒，具有分子伴侣样活性，可在高温下稳定蛋白质。分子伴侣样功能源于合成聚合物纳米颗粒的两个“工程化”性质：1）固有的抗体样蛋白亲和力;和2）低临界溶解温度（LCST），其可用于在升高的温度下捕获蛋白质，防止其聚集和变性，并且在冷却至低于LCST时释放蛋白质而不需要添加剂并且保持蛋白质在溶液中的整体功能。创建“智能”合成聚合物分子伴侣的概念，其中合成聚合物纳米颗粒的抗体样蛋白亲和力用于通过抑制变性和/或蛋白质-蛋白质聚集以及所导致的活性损失来稳定靶蛋白。合成聚合物纳米颗粒的第二个“智能”功能源于其对温度的热响应。该聚合物在高于其较低临界溶解温度的温度下处于塌陷状态，在该温度下其具有蛋白质结合亲和力。这两种独特的聚合物特性允许工程化纳米颗粒的蛋白质结合亲和力，使得其响应于温度而打开和关闭。一种应用是纳米颗粒，其在升高的温度下螯合和稳定靶蛋白，其中蛋白质是脆弱的，但在室温下对蛋白质几乎没有或没有亲和力，以便不妨碍蛋白质功能。具有这种特性的“智能”材料将通过保护蛋白质免于变性而得到应用。它还可以帮助降低蛋白质治疗和诊断的成本，这部分是由于药物在运输和储存过程中的功效保留。该提案将使研究生，本科生和高中生参与跨学科研究计划，涵盖聚合物合成，材料化学，生物学和生物技术。此外，该项目还将为学生提供与工业合作者合作解决实际问题的机会，这可能对全球医疗保健的提供产生深远的影响。蛋白质是由所有生命系统产生的大型生物聚合物。它们在所有生物过程中发挥重要作用，包括调节过程，免疫系统，催化所有生化反应以及其他功能和特性。尽管它们很复杂，但分子生物学和生物技术的进步使蛋白质很容易获得。这些“工程”蛋白质现在被大规模生产和使用，其应用远远超出了其预期的生物功能。这些应用包括治疗和诊断，以及增值化学品的合成。然而，蛋白质有几个限制。特别是，它们在高于室温的条件下通常不稳定，并且这种温度敏感性可以限制蛋白质的使用，特别是在不容易获得制冷设施的情况下。维持蛋白质功能取决于冷藏设施网络的可用性，这些设施将在运输，储存和处理过程中提供最佳的低温。自然界已经进化出了保护蛋白质免受高温影响的策略。一类统称为热休克蛋白的生物大分子，“保护”脆弱的蛋白质在高温下免受功能丧失。这项拟议的研究是开发简单，低成本的合成聚合物，可以模仿热休克蛋白的行为。这些“工程化”合成聚合物纳米颗粒被设计成通过隔离蛋白质来“保护”蛋白质在高温下不变性。该提案将在跨学科研究计划中为从研究生到高中水平的学生提供教育和培训机会，该计划涵盖许多科学领域，如聚合物合成，材料化学，生物技术和生物学。此外，该项目将为学生提供与工业合作者合作的机会，并接受培训，不仅为他们培养下一代科学家，还为他们提供在工业中工作的机会。