CAREER: Nature-Inspired Strategies for Protein Stabilization

职业：受自然启发的蛋白质稳定策略

• 批准号: 1945521
• 负责人: Sarah Perry
• 金额: $ 65.79万
• 依托单位: University of Massachusetts Amherst
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1945521&HistoricalAwards=false
• 关键词: CAREER; Nature; Inspired; Strategies; Protein

NON-TECHNICAL SUMMARYProteins and enzymes perform a wide range of functions inside cells and tissues for long periods of time, and are able to do so at body temperatures. However, most purified proteins used in scientific endeavors, or are present as vaccines or other medical treatments are extremely sensitive to temperature, and must be kept refrigerated. This need to maintain refrigeration during the storage and transportation of these materials represents a significant expense and logistical challenge for both doctors and patients. This proposal will study how different materials can be used to encapsulate such therapeutic proteins to improve their shelf life and temperature sensitivity. This work will also use this funding to create new educational opportunities for students in the lab, and help to develop science-based activities that students at the university could bring home to inspire scientific curiosity in their communities.TECHNICAL SUMMARY This proposal draws upon inspiration from nature, where liquid-liquid phase separated granules in cells are able to selectively sequester specific proteins and other biomolecules, and hold these targets in storage during times of stress. This work will use sequence-controlled polypeptide based complex coacervates as a synthetically accessible mimic for these biological condensates. The effect of charge sequence, hydrophobicity, and hydrogen bonding on the phase behavior of the resulting coacervates will be studied. Furthermore, the effect of these various parameters on the ability of complex coacervate phases to uptake different members of the caspase family of apoptotic enzymes will be evaluated, and correlated with the potential for the different coacervate environments to improve the thermal stability and shelf life of these enzymes. The goal of this effort is to enable the development of general strategies for engineering biomimetic materials for applications ranging from biosensors to thermostable vaccines. Additionally, this proposal will increase educational opportunities at the interface of biology, materials, and engineering at the undergraduate and graduate levels and engage female and minority students through outreach activities with the college's Diversity Programs Office. A new “Innovation in a Box” program will challenge undergraduate outreach leaders to develop easily translatable modules for use in the community.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

非技术概要蛋白质和酶在细胞和组织内长期发挥广泛的功能，并且能够在体温下发挥作用。然而，大多数用于科学研究的纯化蛋白质，或者作为疫苗或其他医学治疗存在的蛋白质，对温度极其敏感，必须冷藏。在这些材料的储存和运输过程中需要保持冷藏，这对医生和患者来说都是巨大的费用和后勤挑战。该提案将研究如何使用不同的材料来封装此类治疗性蛋白质，以提高其保质期和温度敏感性。这项工作还将利用这笔资金为实验室的学生创造新的教育机会，并帮助开展基于科学的活动，大学学生可以将这些活动带回家，以激发社区中的科学好奇心。 技术摘要 该提案汲取了来自大自然的灵感，细胞中的液-液相分离颗粒能够选择性地隔离特定的蛋白质和其他生物分子，并在压力时期将这些目标储存起来。这项工作将使用基于序列控制的多肽的复合凝聚层作为这些生物凝聚物的合成可接近的模拟物。将研究电荷顺序、疏水性和氢键对所得凝聚层的相行为的影响。此外，将评估这些不同参数对复杂凝聚层相吸收凋亡酶半胱天冬酶家族不同成员的能力的影响，并将其与不同凝聚层环境改善这些酶的热稳定性和保质期的潜力相关联。这项工作的目标是开发仿生材料工程的通用策略，其应用范围从生物传感器到热稳定疫苗。 此外，该提案将增加本科生和研究生在生物学、材料和工程领域的教育机会，并通过学院多元化项目办公室的外展活动吸引女性和少数族裔学生。新的“创新盒子”计划将挑战本科生外展领导者开发易于翻译的模块以供社区使用。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Incorporation of Proteins into Complex Coacervates

将蛋白质掺入复杂凝聚层中

• DOI: 10.1016/bs.mie.2020.06.006
• 发表时间: 2021
• 期刊: Methods in enzymology
• 作者: Blocher McTigue, W.C.;Perry, S.L.
• 通讯作者: Perry, S.L.