Design and Evolution of SnoopLigase for Unbreakable Biomolecular Connections

用于牢不可破的生物分子连接的 SnoopLigase 的设计和进化

• 批准号: BB/S007369/1
• 负责人: Mark Howarth
• 金额: $ 49.59万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS007369%2F1
• 关键词: Design; Evolution; SnoopLigase; Unbreakable; Biomolecular

Vaccination is one of the most effective ways to assure the health of humans and other animals and to ensure the security of the food supply. There is an urgent need to improve the way that vaccines are generated, because we lack effective vaccines to many of the most challenging diseases. Also, standard approaches to generate vaccines are slow compared to the speed that outbreaks of new diseases can occur. New vaccines are often developed from Virus-like particles (VLPs), which resemble viruses on the outside, but cannot cause disease because they do not have the virus genome inside. VLPs can be engineered to display a protein from a human or veterinary pathogen on their surface, to create an effective and safe vaccine. Decorating VLPs with such proteins from pathogens is a central problem for vaccine development. VLP decoration requires expensive trial-and-error experiments and has proved unfeasible with many protein targets. We have recently engineered a "bacterial superglue", termed SnoopLigase, which can catalyse the unbreakable linkage of one protein to another protein. This attachment is irreversible and applicable to a wide range of protein targets. SnoopLigase has proved useful in making enzymes more tolerant to harsh conditions and is likely to have diverse applications, including in assembly of diagnostic devices. In this proposal we will use evolution and computer-based design to engineer the next generation of SnoopLigase technology, so that it is faster, active in living cells (not just in the test-tube), and more potent at VLP decoration. SnoopLigase should be a general resource for scientists worldwide and speed up the creation of effective vaccines against major disease challenges.

疫苗接种是确保人类和其他动物健康以及确保食品供应安全的最有效方法之一。迫切需要改进疫苗的生产方式，因为我们缺乏针对许多最具挑战性疾病的有效疫苗。此外，与新疾病爆发的速度相比，生产疫苗的标准方法是缓慢的。新的疫苗通常是从病毒样颗粒（VLP）中开发出来的，VLP在外部类似于病毒，但不会引起疾病，因为它们内部没有病毒基因组。VLP可以被改造成在其表面展示来自人类或兽医病原体的蛋白质，以产生有效和安全的疫苗。用来自病原体的这种蛋白质修饰VLP是疫苗开发的核心问题。VLP修饰需要昂贵的试错实验，并且已被证明对许多蛋白质靶标不可行。我们最近设计了一种“细菌强力胶”，称为SnoopLigase，它可以催化一种蛋白质与另一种蛋白质的牢不可破的连接。这种连接是不可逆的，适用于广泛的蛋白质靶点。SnoopLigase已被证明可用于使酶对恶劣条件更具耐受性，并且可能具有多种应用，包括诊断设备的组装。在这项提案中，我们将使用进化和基于计算机的设计来设计下一代SnoopLigase技术，使其更快，在活细胞中（不仅仅是在试管中）更活跃，并且在VLP装饰中更有效。SnoopLigase应该成为全世界科学家的通用资源，并加速针对重大疾病挑战的有效疫苗的创建。

项目成果

SnoopLigase-Mediated Peptide-Peptide Conjugation and Purification.

SnoopLigase 介导的肽-肽缀合和纯化。

• DOI: 10.1007/978-1-0716-0928-6_2
• 发表时间: 2021
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Buldun CM

DogCatcher allows loop-friendly protein-protein ligation.

• DOI: 10.1016/j.chembiol.2021.07.005
• 发表时间: 2022-02-17
• 期刊: Cell chemical biology
• 影响因子: 8.6
• 作者: Keeble AH;Yadav VK;Ferla MP;Bauer CC;Chuntharpursat-Bon E;Huang J;Bon RS;Howarth M
• 通讯作者: Howarth M

Power to the protein: enhancing and combining activities using the Spy toolbox.

• DOI: 10.1039/d0sc01878c
• 发表时间: 2020-07-28
• 期刊: Chemical science
• 影响因子: 8.4
• 作者: Keeble AH;Howarth M
• 通讯作者: Howarth M

Design and Evolution of Enhanced Peptide-Peptide Ligation for Modular Transglutaminase Assembly.

• DOI: 10.1021/acs.bioconjchem.3c00122
• 发表时间: 2023-06-21
• 期刊: BIOCONJUGATE CHEMISTRY
• 影响因子: 4.7
• 作者: Keeble, Anthony H.;Wood, Dominic P.;Howarth, Mark
• 通讯作者: Howarth, Mark

SpyMask Enables Combinatorial Assembly of Bispecific Binders

SpyMask 实现双特异性结合剂的组合组装

• DOI: 10.17863/cam.106241
• 发表时间: 2024
• 作者: Howarth M