Metal-Protein Interactions Coordinated by Beryllium Ion
Abstract
The objective of this project is to uncover structure-activity relationships that will lead to an improved understanding of the fundamental chemistry of beryllium ion in metal-protein interactions. This research project will test the hypothesis that Be2+ has the potential to form exceptionally stable coordinate bonds with certain types of peptides and proteins. Aim 1. Characterize the interaction of Be2+ with model compounds, peptides, and GSK3b-related proteins using Isothermal Titration Calorimetry, and identify distinctions between Be2+ interactions and those of Mg2+ and Ca2+. Aim 2. Identify peptide and protein-based biomolecules that coordinate Be2+ selectively and stably, using MicroScale Thermophoresis. This project will quantify the Be2+-binding activity of: (a) simple model compounds, (b) rationally-designed peptides, (c) peptide sequences arising from library screening, (d) the Be2+-binding candidate protein GSK3b, (e) truncated derivatives of GSK3b, and (f) point mutant derivatives of GSK3b. Assessment of binding will include affinity, measured as Kd, and stability, measured as the rate constant koff for the dissociation kinetics. The techniques of ITC (Isothermal Titration Calorimetry) and MST (MicroScale Thermophoresis) will be employed. Wherever possible, the same Be2+-binding molecule will be subject to both ITC and MST so that values from each technique can be compared for cross-corroboration. Solvents for the initial dissolution of model compounds, custom-synthesized peptides, etc. will consist of aqueous buffered solutions with NaCl or KC1 to provide ionic strength; divalent cations will be absent in the initial solutions so that Be2+, Mg2+, or Ca2+ can be added during analysis.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 12, 2017
- Source ID
- W911NF1510216
Entities
People
- Ronald K. Gary
Organizations
- Army Contracting Command
- United States Army
- University of Nevada, Las Vegas