CAREER: Nature-Inspired Strategies for Protein Stabilization

非技术性蛋白质和酶在细胞和组织内长时间发挥广泛的功能，并且能够在体温下发挥作用。然而，大多数用于科学工作、或作为疫苗或其他医疗手段存在的纯化蛋白质对温度极其敏感，必须冷藏。在储存和运输这些材料的过程中需要保持冷藏，这对医生和患者来说都是一个巨大的费用和后勤挑战。这项提案将研究如何使用不同的材料来包裹这种治疗性蛋白质，以提高它们的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，并帮助开发基于科学的活动，大学的学生可以将这些活动带回家，在他们所在的社区激发科学好奇心。技术摘要这项提议借鉴了大自然的灵感，细胞中的液-液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。这项工作将使用序列控制的基于多肽的复合凝聚体作为这些生物凝集物的可合成的模拟物。将研究电荷顺序、疏水性和氢键对所得凝聚体的相行为的影响。此外，这些不同的参数对复杂的凝聚相吸收caspase家族的不同成员的能力的影响将被评估，并与不同的凝聚环境改善这些酶的热稳定性和保质期的潜力相关联。这一努力的目标是为从生物传感器到耐热疫苗等各种应用开发工程仿生材料的一般战略。此外，这项提议将增加本科生和研究生在生物、材料和工程学方面的教育机会，并通过与学院多样性项目办公室的外联活动吸引女性和少数族裔学生。一项新的“盒子里的创新”计划将挑战本科生外展领袖开发易于翻译的模块，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势和更广泛的影响审查标准进行评估，被认为值得支持。

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.