Sculpting Dynamic Amphiphilic Structures

生物膜是大多数生物功能的核心，脂质膜越来越多地在生物技术和纳米医学中找到广泛的新应用。这种自组装的两亲性界面可以采用令人惊讶的复杂形状和液晶结构，这些形状和液晶结构在1、2或3维中有序，长度范围从2 - 3纳米延伸到微米。在分子水平上了解界面结构，有序，动力学和微观力学如何取决于化学结构和组成，以及热力学变量，如温度，水合作用和压力，是学习如何操纵这种自组装软界面以创造新颖和有用的结构和新技术的关键，这是本计划的主要目的。我们已经确定了基础科学面临的三个关键挑战：1）不对称; 2）模式化; 3）曲率，长程组织和对称性。我们认为这些挑战的四个主要方面至关重要：i）组成不对称性和双层中两亲物翻转的动力学; ii）横向分离，线张力和微区形成; iii）膜曲率和曲率弹性; iv）脂质头基之间的电荷和偶极相互作用。此外，这四个方面之间存在着复杂的耦合关系，这也是我们关注的重点。我们组建了一个由五个领先的英国大学研究小组组成的团队，涵盖化学，物理和生物物理。这些小组具有互补的专业知识，包括实验室和同步加速器时间分辨X射线衍射，中子散射，固态核磁共振，量热法，生物分子力显微镜，朗缪尔槽和微流体技术，线性和非线性光谱学，原子力显微镜，光谱和光学成像，光镊，微观流变学和理论。这些方法将用于解决三个关键基础科学挑战的不同相互关联的方面。我们将通过严格的管理结构，以及五个研究小组之间的定期会议和人员交流，确保有效地翻译和综合所有研究结果。在该计划先前开发的工程规则和技术的基础上，我们将整合早期的工作，开发脂质结构到活性脂质系统中，例如：水中的自封装液滴界面双层网络;确定尺寸的图案化不对称囊泡：将微流体与智能液滴微工具耦合;热梯度中的磷脂相和囊泡。然后我们将利用这些知识开发三个演示系统：i）人工细胞器。模仿生物学的一些显著功能和特性的人造细胞器机器的发展将导致个性化医疗保健的新方法。ii）快速药物-膜结合筛选。一种区室化的快速药物筛选装置将允许平行测量药物与由平行液滴界面双层或囊泡网络阵列形成的许多人工质膜模拟物（PMM）的相互作用。iii）大膜蛋白的立方结晶。学习如何溶胀脂质立方相将释放我们构建具有数十或数百纳米量级的单位细胞尺寸的立方支架的能力，允许掺入大的膜蛋白（> 50 kD），这些药物是制药业的主要药物靶点。在该计划期间，现有的研究人员和更广泛的工业界人士将开发进一步的生物和生物技术应用。和学术合作者，他们将被适当地纳入计划。

Biomembranes lie at the heart of most biological function, and lipid membranes are increasingly finding a wide range of novel applications in biotechnology and nanomedicine. Such self-assembled amphiphilic interfaces can adopt an astonishing range of complex shapes and liquid-crystalline structures ordered in 1, 2 or 3 dimensions, over length scales stretching from 2 - 3 nanometres, to microns. Gaining an understanding at a molecular level of how interface structure, ordering, dynamics and micromechanics depend upon chemical structure and composition, and thermodynamic variables such as temperature, hydration, and pressure, is the key to learning how we can manipulate such self-assembled soft interfaces to create novel and useful structures and new technologies, and this is the main aim of this Programme. We have identified three key underpinning basic science challenges: 1) asymmetry; 2) patterning; 3) curvature, long-range organisation and symmetry. There are four main aspects underlying these challenges which we consider are of crucial importance: i) compositional asymmetry and dynamics of amphiphile flip-flop across bilayers; ii) lateral segregation, line tension and microdomain formation; iii) membrane curvature and curvature elasticity; iv) charge and dipolar interactions between lipid headgroups. Furthermore, there is a complicated coupling between all of these four aspects, and this is where we will focus much of our attention.We have assembled a team of five leading UK University research groups, spanning Chemistry, Physics and Biophysics. The groups have complementary expertise covering laboratory-based and synchrotron time-resolved X-ray diffraction, neutron scattering, solid-state nuclear magnetic resonance, calorimetry, biomolecular force microscopy, Langmuir trough and microfluidics technologies, linear and non-linear spectroscopies, atomic force microscopy, spectroscopic and optical imaging, optical tweezers, microrheology, and theory. These approaches will be used to attack different inter-related aspects of the three key basic science challenges. We will ensure an efficient translation and synthesis of all of the findings, by a tightly- regulated management structure, and by regular meetings and staff exchanges between the five research groups.Building on the engineering rules and technologies developed previously in the programme, we will integrate the earlier work to develop lipid structures into active lipid systems such as: self-encapsulated droplet interface bilayer networks in water; patterned asymmetric vesicles of defined size: coupling microfluidics with smart droplet microtools; phospholipid phases and vesicles in thermal gradients.We will then use this knowledge to develop three demonstration systems:i) Artificial Organelles. The development of artificial organelle machines which mimic some of the remarkable functions and properties of biology will lead to new approaches for personalized healthcare. ii) Rapid drug-membrane binding screen. A compartmentalised, rapid drug screening device will allow parallel measurements of drug interactions with a number of artificial plasma membrane mimics (PMMs) formed by an array of parallel droplet interface bilayer or vesicle networks.iii) In-Cubo Crystallization of Large Membrane Proteins. Learning how to swell lipid cubic phases will unlock our ability to construct cubic scaffolds with unit cell dimensions of the order of tens or hundreds of nanometres, allowing incorporation of large membrane proteins (>50kD), which are major drug targets for the pharmaceutical industry.Further biological and biotechnological applications will be developed during the course of the Programme by the current Investigators and a wider group of industrial and academic collaborators, who will be brought into the Programme as appropriate.