Michael Rankin, Ph.D.

Pharmaceutically valuable natural compounds are created on complex, assembly line-like machinery in bacteria, plants, and fungus. Using a combination of X-ray crystallography, mass spectrometry, and in vitro pathway reconstitution, I investigated the potential for engineering these pathways to create unnatural natural products. I cloned over 80 constructs and devised successful purification strategies for over 50 protein targets. I was the lab's liaison for the GM/CA sector at the Advanced Photon Source, coordinating the lab's access to beamtime for crystallographic studies. This was where I learned to love mentoring students in both computational and wet lab techniques!

Employed phage display to raise nanobodies against pharmaceutically interesting membrane proteins.

Sulfate is an essential micronutrient required for plant survival. After sulfate is taken into the cell from the plant roots, a network of proteins form the sulfate assimilation pathway. To assess protein-protein interactions in this pathway, I designed and cloned multidomain constructs for in vivo split-luciferase experiments in Arabidopsis protoplasts. I also gained experience in recombinant protein expression in tobacco using an Agrobacterium tumafaciens method. I trained two additional undergraduates during my time in the lab.

Peroxisomes require transporter proteins to move Coenzyme A across the organelle membrane. Using Arabidopsis thaliana as a model organism, I generated RNAi constructs to knock down genes that encoded potential transporters. Additionally, I used yeast and cell-free expression systems to produce the putative transporters for liposome reconstitution. Uptake of radiolabeled Coenzyme A across these proteoliposomes was monitored as a proxy of transporter function.