Title: Research In-Progress Report: An ongoing investigation of Drosera capensis chitinase enzymes using structure prediction, molecular dynamics simulation, and experimental validation techniques
Abstract: Present in all three domains of life (bacteria, archaea, eukarya), chitinases are a ubiquitous class of Glycosyl Hydrolase (GH) enzymes, used by a variety of organisms to cleave chitin, a durable substrate consisting of N-acetylglucosamine subunits. Each organism thus utilizes a unique repertoire of chitinases, suited to their specific needs and functions (e.g. defense against fungal pathogens, insects, nutrient uptake). Several plant families within the order Caryophllales (e.g. Droseraceae, Nepenthaceae) are characterized by their ability to consume insects. One particular carnivorous plant species, the Drosera capensis, secretes a sticky mucilage on its leaflet to capture insect prey and simultaneously digest it. With its genome previously sequenced by the Martin lab, I then use this information to discover and characterize GH19 chitinases from the Drosera capensis, while my co-author Megha Unhelkar characterized GH18 chitinases. As a member of the Luo lab, I then proceeded to conduct 100 ns molecular dynamics (MD) simulation to observe binding dynamics, high-frequency oscillatory behavior and overall conformational changes. Using these models, I learn firsthand protein expression optimization techniques from other senior Martin lab members to attempt experimental validation of select D. capensis chitinases. A collaborative effort between the Martin, Luo, and Butts labs, my fellow lab members and I aim to investigate chitinases in the carnivorous plant, Drosera capensis, and their specialized adaptations in relation to carnivory and defense.