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。蛋白质的工程是生产生物制药的一种广泛使用的方法，用于研究活细胞中的过程和复杂材料的合成。蛋白质由氨基酸的长聚合物形成，它们折叠形成复杂的三维形状。当前可以进行的大多数变化都是这些颗粒表面的简单补充。为了使用类比，这些是分子等同于重新粉刷汽车或添加新的轮胎轮辋。可以通过在蛋白质首次进行（有效地离开工厂之前有效地进行蛋白质）进行编辑来对蛋白质的形状和结构进行更大的变化，但最终必须用与原始蛋白质相同的材料制成这些修饰，因此目前不可能对蛋白质进行大量修饰，因此一旦对蛋白质进行了大规模修饰，就可以简单地修饰。要去除成品的元素，折叠蛋白质并在保留功能性蛋白质的同时，用新零件代替它。在这个项目中，我们正在寻求这样做：我们将开发方法来执行相当于将汽车变成可转换或关闭发动机的分子等效物！基于我们实验室重新加入蛋白质部位的一些最近开发的技术，我们将开发一种方法，最终将使我们能够去除一部分蛋白质，并用用不同材料制成的替代零件代替，这些材料无法使用细胞的机械插入。

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