RUI: Extraction, Purification and Characterization of the Interaction Between Cyclotide and Cyclotide Aggregates with Lipid Bilayers

在化学系大分子、超分子和纳米化学(MSN)计划的支持下，乔治亚·格温尼特学院科学与技术学院的奈维尔·福莱姆、阿贾伊·马利亚、西蒙·姆万格拉和赛拉姆·坦吉拉拉提议研究环状化合物的化学、结构和聚集性质。环肽是一种具有独特结构和特殊稳定性的微型蛋白质，被植物用来抵御害虫。环类化合物的修饰能力可以增强它们在抗生素、诊断试剂和环保农药设计中的潜在应用。为了指导具有不同用途的环状化合物的有效设计，研究人员提出了提取和纯化环状化合物的方法，以了解它们的化学和结构性质、自组装以及它们与脂类的相互作用。预计通过提高对科学、技术、工程和数学(STEM)教育的兴趣，并通过获得使用合成过程、分析仪器和计算技术的技能和专业知识，参与其中的学生的竞争力将被增强，以支持依赖创新和研究在佐治亚州蓬勃发展的技术和生命科学公司的生态系统。这项研究的目标是培养一批能够满足日益增长的经济和社会需求的多样化的科学研究人员。本项目旨在阐明涉及环肽的超分子结构的分子细节及其与脂质双层的相互作用。用分子动力学和蒙特卡罗混合模拟方法研究了环肽聚集体的热力学性质。还将实施基于机器学习的算法来提高模拟速度。本研究旨在加深对不同的环肽如何渗透或调节具有不同磷脂组成的脂质双层膜的理解。此外，还将使用原子力显微镜和毛细管电泳法来探索环肽的膜结合、形态和自组装。本研究的目标包括：(I)利用绿色溶剂有效地提取和纯化环肽；(Ii)确定环肽聚集体中引起自组装的形态、分子间和分子内相互作用；(Iii)确定环肽与脂质双层之间的相互作用模式；(Iv)考察环肽作为绿色农药的软材料、递送剂和支架的应用；以及(V)展示了机器学习辅助计算在预测基于环肽的组装中的分子水平相互作用的应用。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

With the support of the Macromolecular, Supramolecular and Nanochemistry (MSN) Program in the Division of Chemistry, Neville Forlemu, Ajay Mallia, Simon Mwongela, and Sairam Tangirala at the School of Science and Technology, Georgia Gwinnett College, propose to investigate the chemical, structural, and aggregation properties of cyclotides. Cyclotides are mini-proteins with a unique structure and exceptional stability used by plants for protection against pests. The ability to modify cyclotides can enhance their potential applications in the design of antibiotics, diagnostic agents, and eco-friendly pesticides. To guide the efficient design of cyclotides with diverse applications, the investigators propose ways to extract and purify cyclotides, to understand their chemical and structural properties, self-assembly, and their interactions with lipids. It is anticipated that by enhancing interest in science, technology, engineering, and mathematics (STEM) education, and by gaining skills and expertise in using synthetic processes, analytical instruments, and computational techniques, the competitiveness of the students involved will be enhanced for the ecosystem of technology and life science companies that depend on innovation and research to thrive in the state of Georgia. It is a goal of the proposed research to generate a diverse pool of scientific researchers capable of meeting ever-growing economical and societal needs.This project aims to elucidate the molecular details of supramolecular structures involving cyclotides and their interactions with lipid bilayers. The thermodynamic properties of cyclotide aggregates will be investigated using hybrid molecular-dynamics and Monte-Carlo simulations. Machine learning-based algorithms to improve simulation speeds will also be implemented. This research seeks to enhance understanding of how diverse cyclotides permeate or modulate lipid bilayer membranes with varying phospholipid composition. In addition, the membrane binding, morphology, and self-assembly of cyclotides will be explored using atomic force microscopy, and capillary electrophoretic measurements. The goals of this research include: (i) utilizing green solvents to efficiently extract and purify cyclotides; (ii) determining of the morphology, inter- and intra-molecular interactions that give rise to self-assembly in cyclotide aggregates; (iii) ascertaining the mode of interactions between cyclotides and lipid bilayers; (iv) examining applications of cyclotides as soft materials, delivery agents, and scaffolds for green pesticides; and (v) demonstrating the application of machine learning-assisted computation to predict the molecular-level interactions in cyclotide-based assemblies.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.