Grant: Research Tools and Instruments

<p>This proposals requests funding for plate reader with capabilities to perform Fluorescent Polarization (FP) assay in 96-well and 384-well format and low-volume 1536-format. The instrument will serve as a Departmental Core Facility for rapid and cost-effective measurement of inhibition and dissociation constants in the interactions between biomolecules (proteins, DNA) and synthetic small molecules. Measurement of these interactions is a central topic of research in Chemical Biology and Organic/Bioorganic Chemistry. These capabilities are critical for the research of all co-applicants on this proposal. Specific applications enabled by this instrument in specific research groups are listed below. Both direct co-applicants and specific end-users in the Department of Chemistry are listed:</p> <p> 1. Research groups of Todd Lowary, Christopher Cairo, John Klassen, David Bundle will use FP-plate reader for measurement of binding between synthetic carbohydrates and their derivatives with carbohydrate binding proteins, enzymes, toxins, and carbohydrate-specific antibodies. <br /> 2. Research groups of Dennis Hall, Ratmir Derda, Christopher Cairo, Florence Williams, and Fred West will use FP plate reader for the validation of bioactivity and binding of novel small-molecule inhibitors for proteins.<br /> 3. Research groups of John Vederas, Ratmir Derda, Florence Williams and Robert Campbell will use this instrument for measurement of binding interactions between peptides, peptide-derived macrocycles, peptide-derived natural products or antibiotics with their protein-based targets.<br /> 4. Research group of Gibbs-Davis will use this instrument for measurement of activity of DNA-based nanomaterials.</p> <p> The instrument will be accessible to all researchers in our department (30 faculty, 213 graduate students: the main users are expected to be the research groups of 7 faculty in the Chemical Biology section). No instrument with FP-capabilities is available in Chemistry or any facilities proximal to the Chemistry Department. Existing plate readers in the Chemistry Department do not have FP capabilities. Other instruments for measurement of protein ligand interactions exist in the department (e.g., calorimetry or surface plasmon resonance), but they are significantly lower throughput than FP-instrument and require significantly larger amount of reagents.&nbsp;The latter requirement hampers multiple research projects because protein and ligands of interest are often not available in these quantities. Measurement of protein ligand interactions by FP will be critical to accelerate the research, publications, and HQP training for every applicant on this proposal. Specific details are outlined in the descriptions below. <br /> <br /> <br /> </p>