Velia Garcia (Chemistry, PI Chazin)
Study and Characterization of Torsin: Potential Therapeutic Target
Torsin belongs to the family of AAA+ (ATPases Associated with various cellular Activities) macromolecular remodeling enzymes proteins that utilize ATP to perform mechanical work, such as protein unfolding and degradation. The physiological function of torsin is unclear, but it has been suggested through previous studies that it remodels nuclear envelope proteins, such as LINC (Linker of Nucleoskeleton and Cytoskeleton) components. The Olivares lab is interested in understanding how the LINC complex senses and uses mechanical forces to affect nuclear membrane position, alter chromosome dynamics, and control gene expression. My research will be focused on how torsin remodels the LINC complex using biochemical methods, single molecule optical trapping, and through the development of small molecule agonists against torsin enzymatic and cellular function. This will help to develop therapeutics for several diseases that have been linked to torsin mutations, such as early-onset torsin dystonia, a hereditary movement disorder that is characterized by repetitive and involuntary muscle contractions.
Laura Hesse (Pathology Microbiology Immunology, PI Skaar)
Zinc acquisition during Acinetobacter baumannii infection
Zinc is an essential nutrient metal that serves as a structural and enzymatic co-factor for organisms in all domains of life. During bacterial infections, vertebrate hosts employ strategies to limit zinc availability, while bacteria have mechanisms for acquiring the nutrient despite host defenses. Acinetobacter baumannii, an important cause of pneumonia, wound and burn infections, and bacteremia in hospital intensive care units, is predicted to encode both an inner and outer membrane zinc acquisition system, but the specific proteins involved and their role in zinc acquisition have not been determined. My research aims to define the zinc acquisition machinery in A. baumanniiand uncover the role of zinc during A. baumannii pathogenesis. This work will fill a gap in knowledge concerning metal uptake in an important human pathogen and could provide a bacterial system to target with novel therapeutics.
Calvin Larson (Chemistry, PI Sulikowski)
Pulmonary Arterial Hypertension
Pulmonary arterial hypertension (PAH) is a fatal vascular disease within the lungs marked by oxidative stress and endothelial cell proliferation. The free radicals generated under oxidative stress conditions initiate the non-enzymatic metabolism of arachidonic acid (AA) leading to a family of compounds known as the isofurans (IsoF’s). This metabolism is non-stereospecific leading to 8 constitutional isomers each consisting of 32 stereoisomers (256 total isomers). The role of these 256 IsoF’s in PAH remains unclear as their biological production is minimal making traditional isolation methods impossible. Therefore, a stereodivergent synthesis from enantiomerically pure starting materials is being employed to produce these compounds with high enantiopurity, in sufficient amounts for biological studies.
Jackie Picache (Chemistry, PI McLean)
Development of the Biomedical Integrated Online Profiler for Real-time Omics (BIOPRO) to Probe Biochemical Unknowns
Many biochemical unknowns have remained elusive due to limitations in contemporary analyses yet have tractable impacts on human health. To investigate these unknowns, we are developing a technology capable of observing and probing multiple uncharacterized phenotypic perturbations in one experiment. This technology, called the Biomedical Integrated Online Profiler for Real-time Omics (BIOPRO), enables acquisition of multi-omic measurements (e.g., genomic, transcriptomic, proteomic, glycomic, lipidomic, exposomic, and other small molecule) in a single experiment. The BIOPRO bridges a high content screening (HCS) microscope to a high content/high resolution mass spectrometer (HRMS) – both of which have individually led to significant discoveries and translational knowledge. Coupling these two technologies into an automated single assay will allow genetic and/or transcriptomic information from HCS experiments to be cataloged with biochemical species in real time using mass spectrometric assays. To address the challenges of coupling these two technologies, we have developed a microfluidic device module designed to aspirate samples from in vitroand in vivosamples. This sample indexed parallel pump relay (SIPPR) allows highly-precise, automatedsample collection based on a trigger event from the HCS microscope and transfers the sample into the online, automated sample preparation module. The automated sample preparation module uses molecular size fractionation which enables comprehensive analysis of small molecule omics as well as proteomics into a single integrated chromatography system.
Madison Wright (Chemistry, PI Plate)
Thyroglobulin as a model to study the Dynamics of Client Protein-Chaperone Interactions During Folding, Trafficking, and Secretion
The fate of proteins is tightly regulated by the proteostasis network, which governs their folding, trafficking, and degradation. After translation, client proteins depend on interactions with chaperones and folding, trafficking, or degradation factors. These proteostasis components help clients fold into their native state, traffic the protein to the correct location, or degrade improperly folded proteins. While many client protein interactions with proteostasis network components are known, the order of engagement and consequences of perturbations to these interactions are not well understood. Thyroglobulin provides a unique model to study these processes, as it is a large globular protein with many posttranslational modifications that must occur during folding, trafficking and secretion.Due to its large globular structure, thyroglobulin is prone to misfolding and spends a large amount of time in the endoplasmic reticulum to reach its native state. There are well over 50 characterized thyroglobulin mutations, many of which are single point mutations, that lead to misfolding, improper secretion, and ultimately hypothyroidism. My long term goal is to develop a method to analyze time-resolved interactions between thyroglobulin and chaperones and other proteostasis network components. By developing time-resolved interactomics approaches using mass spectrometry, Bio-orthogonal Noncanonical Amino Acid Tagging (BONCAT) and other chemical biology tools we will be able to better define these interaction networks. This will ultimately reveal underlying mechanisms that are associated with misfolding of disease variants of thyroglobulin that were previously unseen or not well understood. My interactomics findings will then direct the basis for developing new therapeutic strategies by targeting specific factors involved in mistimed or improper protein-protein interactions within disease states.
Jonah Zarrow (Chemical Physical Biology, PI Davies)
Characterizing and inhibiting the synthesis of gut microbiome-produced fatty acyl signaling molecules
N-acyl amides (NAAs) are a superfamily of bioactive compounds that appear to arise in part from bacterial metabolism of dietary components, and that can alter host physiology. As NAAs have been implicated in diet-induced obesity, type 2 diabetes, endocannabinoid signaling, atherosclerosis, inflammation, bone growth, immune responses, and much more, their study is very important for human health. As NAAs are a diverse group of molecules, the first aim of my project will be to use LCMS to determine which NAAs are made by gut bacteria. This will be the first broad characterization of the NAAs that can be made by gut bacteria, and will establish which NAAs (and their synthetic enzymes) are productive avenues for future research. Another important way of studying NAAs is to make probe compounds to modulate their activities. Much research has been done on N-acyl ethanolamides and one of their synthesizing enzymes, NAPE-PLD, because they have a role in obesity, satiety, inflammation, and diabetes. However, there is a lack of potent and selective modulators of NAPE-PLD activity. Therefore, the second aim of my project is to establish structure-activity relationships and use medicinal chemistry to make recently obtained high throughput screen hits of NAPE-PLD inhibitors more potent. Along similar lines, my third aim is to use genomic information to identify enzymes that synthesize N-acyl-oxyacyl amino acids (NAOAs)—which are biosynthesized by joining an NAA to an acyl chain—and then to use HTS to identify compounds that inhibit those enzymes. NAOAs have been shown to have inflammatory, hepatotoxic, and atherosclerotic effects, so inhibitors of their synthesis will both help research efforts and have the potential to be therapeutically useful.
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