During this summer I worked with Dr. Gaynor Wild, Associate Professor in Biochemistry at UNM School of Medicine, and Ron Hughen, a (then) senior undergraduate. One of our two projects was to isolate a glycolipid from brains in mice affected by the Niemann-Pick, Type-C, disease. Niemann-Pick disease is a genetic disorder in which neurotransmitter reuptake is inefficient, due to defective glycolipid structure in neuronal membranes. We obtained quantitative results that showed remarkable difference in concentration of a certain glycolipid between the affected and normal brains.
In another project, we examined the enzyme kinetics of Neuron Specific Enolase (NSE). After simulating in vivo conditions in vitro, we used Hughen's protocol to measure product development spectrophotometrically. We analyzed the data in several ways, including Michaelis-Menten and Eadie-Hofstee analyses. NSE proved to be allosterically regulated.
I joined the Krause Group at the end of my sophomore year with a growing interest in understanding molecular structure. By doing x-ray crystallography and using graphical simulation, I would gain insight to principles of macromolecular structure, methods in structure determination and molecular modeling. This has served as an excellent introduction to application of these sciences to discovery in molecular medicine.
Triple helix DNA technology is a conceptually ideal method of regulating gene expression, at the level of transcription. Certain oligonucleotides have proven to engage genomic DNA to produce a stable triple helix. Understanding the exact relationship between the third strand and the duplex is essential in designing an appropriate oligonucleotide to target a gene coding for a protein culpable for a particular disease. Once the triplex is formed, mRNA can not be synthesized, as mRNA polymerase will not associate properly with the triplex; thus the defunct protein is never produced.
I have crystallized several oligonucleotides unique in sequence and solubility. Having performed preliminary x-ray diffraction analyses on them, we have directed our research into creating higher order crystals of triple helices.
This summer, two graduate students and I formed the Nuclease Group. Our aim is to accurately describe the docking of inhibitors to Serratia Nuclease to allow insight to the mechanism of nuclease assisted hydrolysis of nucleic acids. We are studying nuclease crystals soaked in solutions that contain inhibitors which may form a stable protein-inhibitor complex. Using x-ray diffraction and modeling/dynamics methods, we hope to reveal the non-covalent interactions involved in substrate docking, as well as to identify the catalytically active residues.
Both apolipoprotein and Rad51 are undergoing crystallization experiments.
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