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

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.