The project goal is to understand the transition metal binding specificity of metalloregulatory proteins. The practical applications of this endeavor are at least two-fold: 1) several metalloregulatory proteins currently under investigation come from antibiotic-resistant pathogenic bacteria. Since no human analogue of these regulators exist, targeting them for drug therapy offers an attractive alternative to current antibiotic regimes. 2) Approximately 70% of the proteins in the human body bind metals. A firm understanding of the metal-binding site and hence the selection mechanism will be a huge first step in understanding the function of all metallo-proteins. To this end, we have chosen the Mn2+-dependent regulator MntR from Bacillus subtilis as our model of study. With metal-free starting material in hand, biochemical and biophysical characterization will proceed. The biochemical will focus on the native structural state of the protein (monomer/dimmer), metal-residue contacts, and protein/DNA interaction to name a few. The biophysical will include probing the metal-binding site with Electron Paramagnetic Resonance (EPR) spectroscopy, measuring the binding affinity of the regulator to DNA using Isothermal Titration Calorimetry (ITC) and studying the solution structure of the protein using Analytical Uitracentrifugation. With these tools and more at our disposal, we are confident that a more thorough understanding of the transition metal switch will be found.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31GM069332-04
Application #
7088805
Study Section
Minority Programs Review Committee (MPRC)
Program Officer
Toliver, Adolphus
Project Start
2003-07-28
Project End
2008-07-27
Budget Start
2006-07-28
Budget End
2007-07-27
Support Year
4
Fiscal Year
2006
Total Cost
$28,540
Indirect Cost
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093