Design and Evolution of SnoopLigase for Unbreakable Biomolecular Connections

接种疫苗是确保人类和其他动物健康和确保粮食供应安全的最有效方法之一。迫切需要改进疫苗的产生方式，因为我们缺乏针对许多最具挑战性的疾病的有效疫苗。此外，与新疾病爆发的速度相比，生产疫苗的标准方法很慢。新疫苗通常是从病毒样颗粒(VLP)开发出来的，这种颗粒在外表上与病毒相似，但不会导致疾病，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，以创造出有效和安全的疫苗。用病原体的这种蛋白修饰VLP是疫苗开发的核心问题。VLP的修饰需要昂贵的反复试验，事实证明，对于许多蛋白质靶标来说，这是不可行的。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之间牢不可破的连接。这种结合是不可逆的，适用于广泛的蛋白质靶标。事实证明，SnoopLigase有助于提高酶对恶劣条件的耐受性，并可能有多种应用，包括在诊断设备的组装中。在这个方案中，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，以便它更快，在活细胞中(不仅仅是在试管中)活跃，并在VLP装饰方面更有效。SnoopLigase应该成为世界各地科学家的通用资源，并加快创造有效的疫苗来应对重大疾病挑战。

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.