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

反转开放阅读框以创建生活的伪镜像

• 批准号: 2218373
• 负责人: Christophe Herman
• 金额: $ 30万
• 依托单位: Baylor College of Medicine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2218373&HistoricalAwards=false
• 关键词: Reversing; open; reading; frames; create

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.

这个项目探讨了逆转蛋白质中氨基酸顺序的后果。逆转蛋白质的合成顺序有可能发现增强的或全新的蛋白质功能，从而帮助改善日常生活。加速化学反应的蛋白质通常用于洗涤剂，牙膏和燃料处理，仅举几个家庭用品。这些活性蛋白质取自自然界，可以改进以更好地发挥作用。然而，随着每一次改进，进一步的改进变得更加难以实现。逆转一种蛋白质以一种新的方式重新创造了一种天然蛋白质，这种蛋白质可能能够突破这些障碍，以进一步改进。对这些反向蛋白质的研究正在与包括科学中代表性不足的群体成员在内的学生进行，以便下一代科学家将准备好用这些活性蛋白质改善我们的生活。由于新的生命形式的合成开辟了几条生物伦理学调查的路线，这些问题首先在小组内得到解决，然后更广泛地通过学术生物伦理学报告和公众通过在当地博物馆举办的一系列研讨会得到解决。 为了制造具有功能的反向蛋白，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正确折叠的氨基酸残基。这些鉴定的氨基酸正在计算机上改变，以确定该位置的不同氨基酸是否改善折叠。这些计算预测也正在进行实验测试，以确定它们是否改善了反向蛋白质的折叠和功能。在一个平行的方法中，反向蛋白质的功能正在通过定向进化得到改善。这些改进的反向蛋白质正在纯化和测试体外功能，热稳定性和蛋白质结晶的适用性，以便可以确定它们的3D结构。这些实验的发现提高了对蛋白质折叠和功能的理解，并使科学家能够创造具有增强功能或新功能的新蛋白质。该奖项反映了NSF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。