Elizabeth Clark
Pronouns: she/her/hers
Research Mentor(s): Janet Smith
Research Mentor School/College/Department: Life Sciences Institute / Medicine
Program:
Authors: Elizabeth Clark, Jamie Konwerski
Session: Session 3: 11:00 am – 11: 50 am
Poster: 28
Abstract
Polyketide synthases (PKS) and PKS-like synthases are enzymatic protein complexes that play a large role in the biosynthesis of a wide variety of complex, therapeutically relevant compounds. These unique pathways exhibit meaningful anticancer, antibiotic, and anesthetic properties, with strong potential for the advancement of many different fields such as drug discovery, medicine, or agriculture. One such compound, saxitoxin, is a potent neurotoxin discovered in various cyanobacteria. My project will utilize the PKS-like megasynsthase module SxtA, from the cyanobacteria Microseira wollei. The final product of this module is the starting unit for saxitoxin. SxtA is a unique four-domained “assembly line” module comprised of a MT (methyltransferase), DC (decarboxylase), ACP (acyl carrier protein), and an AONS (8-amino-7-oxononanoate synthase) domain. I will investigate one of four available constructs for the AONS domain in the SxtA module. To optimize purification of the AONS domain, I will utilize PCR for recombinant gene optimization, site-directed mutagenesis to generate active site mutations, E. coli for protein expression, and affinity and size exclusion chromatography for protein purification. X-ray crystallography techniques will be used to determine the three-dimensional structure of the protein. As a result, I expect to achieve a solved structure of the purified AONS domain through protein crystallization, which is in line with my project’s goal. This information can then be applied by other researchers and scientists who are trying to understand the chemical mechanism and overall biological effects of these complex pathways.
