Enhancing the innate cellular degradation capacity

摘要：蛋白质的生物发生涉及多肽链通过一系列折叠中间体的进展，以寻找最热动力学稳定的构象-天然3D结构。发生在拥挤的细胞环境中，这一过程可能导致原生折叠或偏离途径的错误折叠物种。蛋白质错误折叠不仅会损害细胞功能，还会有利于聚集体的形成，从而导致细胞的破坏性后果。在真核细胞中，折叠质量控制系统平衡蛋白质的合成、折叠和降解，从而维持蛋白质的稳态。操纵蛋白质表达以增加蛋白质生产扰乱蛋白质稳态，导致错误折叠蛋白质的积累和聚集体的形成。错误折叠和聚集反过来导致诱导蛋白毒性应激和细胞凋亡的激活。错误折叠蛋白质的积累有利于新合成蛋白质的错误折叠，进一步损害蛋白质组的折叠和稳定性，并且聚集体的形成具有高度的细胞毒性。毫不奇怪，蛋白质错误折叠和聚集是重组蛋白高产生产的关键障碍，重组蛋白是制药行业增长最快的领域之一。调控错误折叠蛋白处理的分子机制仍然难以捉摸。该项目将解决这些基本的细胞生物学问题，并将利用合成生物学方法和蛋白质工程技术来破译和操纵先天的细胞降解能力。这项工作的结果将有助于设计新的细胞工程策略，以限制错误折叠蛋白质的积累，防止聚集并提高天然折叠蛋白质的产生。这些策略有可能显著影响用于研究、工业和诊断应用的重组蛋白的高产生产。此外，从这项研究中获得的知识将促进我们对衰老和蛋白质错误折叠疾病的理解。提出的实验计划将提供一个教育本科生的环境。它还将创造一个有效的平台，以接触高中生，激发他们对科学和技术的兴趣，目标是增加和多样化对生物技术相关学科感兴趣的学生群体。教育计划包括创建一系列指导项目和研究经验，吸引本科生和高中生，特别强调妇女和未被充分代表的少数民族。

Abstract#1159640Segatori, LauraProtein biogenesis involves the progression of a polypeptide chain through a series of folding intermediates in search for the most thermodynamically stable conformation - the native 3D structure. Taking place in the crowded cellular milieu, this process can lead to native folding or off-pathway misfolded species. Protein misfolding can result in destructive consequences for the cell as it not only impairs functions but can also favor formation of aggregates. A folding quality control system balances synthesis, folding and degradation of proteins thereby maintaining protein homeostasis in eukaryotic cell. Manipulating protein expression to increase protein production perturbs protein homeostasis and results in accumulation of misfolded proteins and formation of aggregates. Misfolding and aggregation, in turn, cause induction of proteotoxic stress and activation of apoptosis. The accumulation of misfolded proteins favor misfolding of newly synthesized proteins, further compromising folding and stability of the proteome, and the formation of aggregates is highly cytotoxic. Not surprisingly, protein misfolding and aggregation are key impediments to the high-yield production of recombinant proteins, which is one of the fastest growing sectors of the pharmaceutical industry. The molecular mechanisms that regulate the disposal of misfolded proteins still remain elusive. This project will address these fundamental cell biology questions and will make use of synthetic biology approaches and protein engineering techniques to decipher and manipulate the innate cellular degradation capacity. Results from this work will enable the design of novel cell engineering strategies to limit accumulation of misfolded proteins, prevent aggregation and enhance production of natively folded proteins. These strategies have the potential to significantly impact the high yield production of recombinant proteins for research, industrial and diagnostic applications. In addition, the knowledge gained from this study will advance our understanding of aging and protein misfolding diseases.The proposed experimental plan will provide a setting to educate undergraduate students. It will also create an effective platform for reaching out to high school students and inspire their interest in science and technology with the goal to increase and diversify the pool of students interested in biotechnology related disciplines. The educational plans incudes creating a series of mentoring programs and research experiences to engage undergraduate and high school students, with particular emphasis to women and underrepresented minorities.