CAREER: Does Crowding Stabilize Intrinsically Disordered Proteins?

AbstractIntellectual优点。蛋白质是复杂的生物分子，几乎调节着每一个细胞过程。一个长期存在的范式是，蛋白质的结构，即其原子的三维排列，决定了蛋白质的功能。然而，过去十年的研究已经发现了大量的内在无序蛋白（IDPs），它们通过没有明确定义的三维结构的灵活区域来实现其功能。对IDPs重要性的日益认识正在推翻长期以来关于蛋白质结构和蛋白质功能之间联系的观点。在细胞中，蛋白质分子紧密地聚集在一起，导致一种被称为拥挤的现象。众所周知，拥挤会影响结构蛋白，但对拥挤对内部流离失所者的影响知之甚少。虽然很明显，在原始的实验室条件下，IDPs缺乏结构，但在细胞内高度拥挤的环境中，这些蛋白质是否保持无结构尚不清楚。因此，在了解拥挤对国内流离失所者的影响方面作出了相当大的努力。这些努力包括在实验室中模拟拥挤，以及在细胞中直接探测IDPs。尽管取得了进展，但获取拥挤条件下国内流离失所者结构的信息仍然具有挑战性。在应对这一挑战之前，对IDP功能的理解将是有限的。因此，这个项目的目标是了解拥挤如何影响国内流离失所者的结构和灵活性。在这个项目中，将确定拥挤对一组国内流离失所者的影响。预期的结果将是：(1)对拥挤对国内流离失所者影响的新见解；(2)可用于获取拥挤条件下其他国内流离失所者行为信息的变革性方法。这项研究的成功完成将导致对内在无序蛋白质如何在细胞中起作用的更深层次的理解。更广泛的影响。该项目将通过吸引本科生和研究生回答有关蛋白质结构的基本重要问题来促进发现，同时促进教学和学习。该提案的教育目标是通过开发一门课程来提高科学素养，培养未来的科学教师，并与当地社区分享知识，从而使分子结构原理及其对生物学的影响得到广泛传播。通过其教育目标，本项目将通过向本科生和公众广泛传播分子结构原理，通过发展对这些原理如何应用于现代生活的理解，通过支持未来中学科学教师的发展，以及通过增加对科学研究的理解，产生重要的社会效益。最后，这项研究将通过支持代表性不足的群体在科学领域的参与，扩大代表性不足群体的参与。

AbstractIntellectual merit. Proteins are complex biomolecules that regulate nearly every cellular process. A long-standing paradigm is that the structure of a protein, that is, the three-dimensional arrangement of its atoms, determines how the protein functions. Research over the past decade, however, has uncovered a vast universe of intrinsically disordered proteins (IDPs) that carry out their functions through flexible regions that do not have a well-defined three-dimensional structure. The growing recognition of the importance of IDPs is overturning long-held ideas about the connections between protein structure and protein function. In cells, protein molecules are packed tightly together leading to a phenomenon known as crowding. Crowding is known to affect structured proteins, but much less is known about the effects of crowding on IDPs. While it is clear that IDPs lack structure under pristine laboratory conditions, it is not at all clear whether such proteins remain unstructured in highly crowded environments found inside cells. Thus considerable effort has been directed at understanding the effects of crowding on IDPs. Such efforts include attempts to simulate crowding in the laboratory and efforts to probe IDPs directly in cells. Despite advances, obtaining information about IDP structure under crowded conditions remains challenging. Until this challenge is met, understanding of IDP function will be limited. Therefore, the goal of this project is to understand how crowding affects the structure and flexibility of IDPs. In this project, the effects of crowding on a panel of IDPs will be determined. The expected outcomes will be (1) new insight into the effects of crowding on IDPs and (2) transformative methodology that can be used to obtain information about the behavior of other IDPs under crowded conditions. Successful completion of this research will lead to a deeper understanding of how intrinsically disordered proteins function in the cell. Broader impacts. This project will advance discovery while promoting teaching and learning by engaging both undergraduates and graduate students to answer fundamentally important questions about protein structure. The educational objective of this proposal is to enable broad dissemination of the principles of molecular structure and their impact on biology, by developing a course that increases scientific literacy, develops future science teachers, and shares knowledge with the local community. Through its educational objective, this project will have important societal benefits by broadly disseminating the principles of molecular structure to undergraduate students and the general public, by developing an understanding of how these principles apply to modern life, by supporting the development of future secondary science teachers, and by increasing understanding of scientific research. Finally, this research will broaden participation of underrepresented groups by supporting underrepresented groups in science.