Rational engineering of self-assembling thermo-sensitive protein complexes and their applications

这一工业生物技术项目依赖于生物资源和材料，如SNARE蛋白激发的分子自组装系统，用于生产热敏性药物前体、药物载体和热敏纳米颗粒。自然产生的SNAR蛋白驱动每个真核细胞的细胞膜融合。利用这些，我们开发了一种新的分子装订方法，允许重组蛋白的定位、非共价自组装(WO2011/018665A1)。这种合理设计的超分子自组装系统优于所有已知的蛋白质自组装系统，这是因为它对杂色剂、强洗涤剂、蛋白酶、高温以及其穿透细胞的特性具有抵抗力。然而，诱捕器用于药物输送因其干扰本地诱捕器和胞吐过程的能力而受到阻碍。我们的研究努力旨在解决这个遗留问题。这项研究的总体目标是设计一种更广泛的圈套诱导蛋白，这些蛋白能够控制自组装和拆解，并且不干扰天然的圈套，因此适合在体内应用。有两个项目可供选择。该项目的一个目标是确定扩大范围的工程诱捕蛋白的关键特征，这些特征是诱捕蛋白具有卓越的结构稳定性的原因。我们的方法包括分子建模、计算分析、重组蛋白表达、生化、生物物理、结构和热分析。我们还将测试分子货物对这些蛋白质复合体的自组装速度和整体稳定性的影响。该项目是米哈伊尔·索洛维耶夫(伦敦皇家霍洛威大学)、恩里科·法拉利(林肯大学)和朱利亚诺·西里加迪(钻石光源，哈威尔科学校区)共同开展的研究。研究将在所有三个地点进行。该项目的另一个目标是设计基于蛋白质的纳米颗粒，用于靶向药物输送。我们的方法包括分子建模、计算分析、重组蛋白表达、生化和热分析。我们还将测试这些纳米颗粒与一系列脂类的兼容性，以及它们对我们的组合生物微粒系统热稳定性的影响。这个项目是米哈伊尔·索洛维耶夫(伦敦皇家霍洛威大学)和萧耘·徐(伦敦帝国理工学院)的合作研究。

This industrial biotechnology project relies on the biological resources and materials such as SNARE proteins-inspired molecular self-assembling system for producing thermosensitive pharmaceutical precursors, drug carriers and theranostic nanoparticles. Naturally occurring SNARE proteins drive fusion of cellular membranes in every eukaryotic cell. Using these we have developed a new molecular stapling method which allows site-oriented, non-covalent self-assembly of recombinant proteins (WO2011/018665A1). Such rationally engineered supramolecular self-assembling system is superior to all protein self-assembling systems known due to its resistance to chaotropic agents, strong detergents, proteases, elevated temperatures as well as its cell-penetrating properties. However the use of SNARES for drug delivery is hindered by their ability to interfere with native SNARES and the process of exocytosis. Our research effort is aimed at resolving this remaining problem. The overall aim of this research is engineer an expanded range of SNARE-inspired proteins which are capable of controlled self-assembly and dis-assembly and which do not interfere with native SNARES and are therefore suitable for in vivo applications. Two projects are available. One objective of this project is to identify key features of the expanded range of engineered SNARE-inspired proteins, responsible for the superior structural stability of SNAREs. Our methodology includes molecular modelling, computation analysis, recombinant protein expression, biochemical, biophysical, structural and thermal analyses. We will also test the effect of molecular cargo on the rates of self-assembly and the overall stability of these protein complexes. This project is a collaborative research between Mikhail Soloviev (Royal Holloway University of London), Enrico Ferrari (University of Lincoln) and Giuliano Siligardi (Diamond Light Source, Harwell Science Campus). Research will be conducted at all three sites. Another objective of this project is to engineer protein-based nanoparticles for targeted drug delivery. Our methodology includes molecular modelling, computation analysis, recombinant protein expression, biochemical and thermal analyses. We will also test these nanoparticles for their compatibility with a range of lipids and their effect on the thermal stability properties of our combinatorial bioparticle system. This project is a collaborative research between Mikhail Soloviev (Royal Holloway University of London) and Xiao Yun Xu (Imperial College London).