Reversing open reading frames to create a pseudo-mirror image of life

这个项目探索颠倒蛋白质中氨基酸顺序的后果。颠倒蛋白质的生成顺序有可能发现增强的或全新的蛋白质功能，有助于改善日常生活。加速化学反应的蛋白质通常用于洗涤剂、牙膏和燃料处理，仅举几例。这些活性蛋白质取自大自然，经过改良后可以更好地发挥作用。然而，随着每一次改进，进一步的改进变得更难实现。逆转一种蛋白质以一种新的方式重建了一种天然蛋白质，这可能能够突破这些障碍，进一步改善。对这些反向蛋白质的研究正在与学生一起进行，这些学生包括科学界代表性不足的群体的成员，以便下一代科学家准备好用这些活性蛋白质改善我们的生活。由于新生命形式的合成开辟了生物伦理研究的几条路线，这些问题首先在小组内部得到解决，然后在更广泛的范围内，通过学术生物伦理报告和通过当地博物馆的既定系列研讨会向公众提出。为了制造具有功能的反向蛋白质，贝勒医学院的研究人员将使用机器学习算法AlphaFold2来识别阻止反向蛋白质正确折叠的氨基酸残基。这些已识别的氨基酸在计算机上被改变，以确定在该位置的不同氨基酸是否能改善折叠。这些计算预测也正在进行实验测试，以确定它们是否能改善反向蛋白质的折叠和功能。在一个平行的方法中，反向蛋白的功能正在通过定向进化得到改善。这些改进的反向蛋白正在进行纯化和体外功能、热稳定性和蛋白质结晶性的测试，以便确定它们的3D结构。这些实验的发现正在提高人们对蛋白质折叠和功能的理解，并使科学家能够创造出具有增强功能或新功能的新蛋白质。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This project explores the consequences of reversing the order of amino acids in a protein. Reversing the order in which a protein is made has the potential to discover enhanced or completely new protein functions that can help improve daily life. Proteins that speed-up chemical reactions are routinely used in washing detergents, toothpaste, and fuel treatments, to name a few household items. These active proteins are taken from nature and can be improved to do their job better. However, with each improvement, further improvements become harder to attain. Reversing a protein recreates a natural protein in a new way that may be able to break through these barriers to further improvement. Research on these reverse proteins is being conducted with students that include members of underrepresented groups in science so that the next generation of scientists will be ready to better our lives with these active proteins. As the synthesis of new life forms opens up several lines of bioethical inquiry, these are addressed, first within the group, and then more broadly, with academic bioethics reports and with the general public through an established seminar series at a local museum. To make reverse proteins that function researchers at Baylor College of Medicine will use the machine learning algorithm AlphaFold2 to identify amino acid residues that prevent proper folding of reverse proteins. These identified amino acids are being changed in silico to determine if a different amino acid at that position improves folding. These computational predictions are also being tested experimentally to determine if they improve folding and function of the reverse protein. In a parallel approach, the functions of reverse proteins are being improved through directed evolution. These improved reverse proteins are being purified and tested for in vitro function, thermal stability, and suitability for protein crystallization so that their 3D structure can be determined. The findings from these experiments are improving understanding of how proteins fold and function and allowing scientists to create new proteins with enhanced or new functions.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.