Combining Molecular Dynamics and Neutron Reflectometry Techniques to Understand Lipid Transfer Protein Binding Events at the Membrane Interface

植物脂转移蛋白(PLTP)具有结合和运输脂质的能力，被认为是植物在陆地上生存的关键蛋白。PLTP在植物生长、抵御生物和非生物胁迫、脂质屏障沉积、有性繁殖和信号转导等过程中发挥着重要作用。然而，对PLTPs的结构和功能的研究仍然很少，其生物学作用也仍然是未知的。本项目旨在通过结合中子反射仪(NR)和分子动力学(MD)模拟来研究PLTP与由脂类混合物组成的自由漂浮膜系统之间的相互作用，从而获得对PLTP活性的分子洞察。由于NR是一种非破坏性的方法，使我们能够测量在生理条件下蛋白质添加到膜系统中的情况，因此它是一种获得动态膜结合过程的结构洞察的强大技术。NR实验提供的结构信息可以用来生成MD模拟的起始条件和参数，从而产生相互作用过程中膜和蛋白质结构的完全原子分辨率，从而使我们能够改进实验模型。BBSRC优先领域通过结合中子反射和分子动力学模拟来提高对植物脂转移蛋白的理解的方法解决了几个BBSRC优先领域。特别是，通过使用和开发进一步的计算工具来完善和支持实验中已获得的结构模型的方法涉及到“数据驱动生物学”和“生物科学的系统方法”领域。获得与植物膜相关的现象的精确分子细节对于我们理解植物中广泛的生化过程，如病原体相互作用、细胞动态平衡和调控，具有潜在的巨大的下游益处。因此，该项目的见解可以导致新的植物处理策略，从而提高食品生产的效率。因此，“可持续提高农业生产”这一优先领域也通过该项目得到解决。

Plant lipid transfer proteins (PLTPs) have the ability to bind and transport lipids and are considered as key proteins for plant survival on land. PLTPs play a crucial role in many processes in plants, such as growing, defence against biotic and abiotic stress, lipid barrier deposition, sexual reproduction and signalling. However, the structural and functional investigations of PLTPs are still lacking and their biological role remains elusive. This project aims to gain molecular insights into the activity of PLTPs by combining neutron reflectometry (NR) and molecular dynamic (MD) simulations to study the interaction between PLTPs with free floating membrane systems composed of lipid mixtures which accurately represent the compositions and fluidity of higher plant membranes. NR is a powerful technique for gaining structural insights into dynamic membrane binding processes as it is a nondestructive method which enables us to measure the addition of a protein to a membrane system at physiological conditions. The structural information provided by the NR experiments can be used to generate starting conditions and parameters for MD simulations, which produce a fully atomisticresolution of the membrane and protein structure during the interaction process, thereby enabling us to refine the experimental model.Addressed BBSRC Priority AreasThe approach to improve the understanding of plant lipid transfer proteins through the combination of neutron reflectometry and molecular dynamic simulations addresses several BBSRC priority areas. In particular, the approach to refine and support the experimental achieved structural models by using and developing further computational tools relates to the area of 'data driven biology' and 'systems approaches to the biosciences'.Gaining precise molecular details of plant membrane related phenomena has potential large downstream benefits to our understanding of a wide range of biochemical processes in plants such as pathogen interactions, cell homeostasis and regulation. Thus, the insights of this project can lead to new treatment strategies of plants and therefore improve the efficiency of food production. Hence, the priority area of 'Sustainably enhancing agricultural production' is also addressed through this project.