21ENGBIO - Peptide excision, replacement and ligation (PERL) as a new strategy for protein engineering

21ENGBIO - 肽切除、替换和连接 (PERL) 作为蛋白质工程的新策略

• 批准号: BB/W01131X/1
• 负责人: Michael Webb
• 金额: $ 12.84万
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW01131X%2F1
• 关键词: 21ENGBIO; Peptide; excision; replacement; ligation

In this project, we will develop a completely new way to re-engineer the basic structure of proteins. Engineering of proteins is a widely-used approach for the production of biopharmaceuticals, for the study of processes in the living cell and for the synthesis of complex materials. Proteins are formed of long polymers of amino-acids which fold up to form complex three-dimensional shapes. The majority of changes that can currently be made are simple additions to the surface of these particles. To use an analogy, these are the molecular equivalent of repainting a car or adding new tire rims. Larger changes to the shape and structure of the protein can be made by editing the protein before it is first made (effectively before it leaves the factory) but ultimately these modifications must be made of the same materials as the original protein and so some modifications are simply inaccessible at the moment.What it is not currently possible to do is to make large modifications to the protein once it has been made; to remove an element of the finished, folded protein and replace it with a new part while retaining a functional protein. In this project, we are seeking to do just that: we will develop methods to carry out the molecular equivalent of turning a car into a convertible or switching out the engine! Based on some recently developed technology in our laboratory to rejoin the protein parts, we will develop a method that will ultimately allow us to remove a part of the protein and replace it with alternative parts made from different materials that cannot be inserted using the machinery of the cell.

在这个项目中，我们将开发一种全新的方法来重新设计蛋白质的基本结构。蛋白质工程是一种广泛使用的方法，用于生产生物药物，研究活细胞中的过程和合成复杂材料。蛋白质是由氨基酸的长聚合物形成的，这些聚合物折叠起来形成复杂的三维形状。目前可以进行的大多数改变都是对这些颗粒表面的简单添加。打个比方，这些分子相当于重新粉刷汽车或添加新的轮胎轮辋。蛋白质的形状和结构的更大变化可以通过在蛋白质首次制造之前编辑蛋白质来实现（在其离开工厂之前有效地），但是最终这些修饰必须由与原始蛋白质相同的材料制成，并且因此一些修饰目前根本无法进行。去除最终折叠蛋白质的一个元素，并用新的部分替换它，同时保留功能性蛋白质。在这个项目中，我们正在寻求做到这一点：我们将开发方法来进行分子相当于把一辆汽车变成敞篷车或切换引擎！基于我们实验室最近开发的一些重新连接蛋白质部分的技术，我们将开发一种方法，最终使我们能够去除蛋白质的一部分，并用由不同材料制成的替代部分代替它，这些材料不能使用细胞的机器插入。

项目成果

Quantitative N- or C-Terminal Labelling of Proteins with Unactivated Peptides by Use of Sortases and a d -Aminopeptidase

使用分选酶和 d-氨基肽酶对带有未激活肽的蛋白质进行定量 N 或 C 末端标记

• DOI: 10.1002/ange.202310862
• 发表时间: 2024
• 期刊: Angewandte Chemie
• 作者: Arnott Z