This shared instrumentation grant program proposal requests funds for the purchase of a state-of-the-art isothermal titration microcalorimeter (ITC). Calorimetry provides a wealth of information about macromolecular interactions and enzymatic kinetics by measuring the heats of reaction. This technique has distinct advantages over others, including that it can obtain information about the binding affinity, enthalpy and entropy of reaction and stoichiometry of the reaction in a single experiment and that it does not require the labeling of either the titrant or titrate molecules. Recent advances in instrumentation also allow for 10-fold lower sample amounts to be used per experiment and significantly decreased run time as compared to previous generation instruments. These considerations make this technique accessible to a broader spectrum of users and facilitates overall higher throughput, which are important features for shared instrumentation. Acquisition of the proposed MicroCal iTC200 would facilitate research programs directly related to human health issues in a number of important ways. Major users of this instrument propose to use this instrument to investigate the regulatory mechanisms of kinases involved in a variety of key cellular signal transduction pathways in humans (Prof. Natalie Ahn), RNA-based regulatory elements that directly sense cellular metabolites (Prof. Robert Batey), a nucleic acid aptamer that is an FDA- approved drug to fight macular degeneration (Prof. Arthur Pardi), and the nature of protein-DNA interactions at the end of linear chromosomes (Prof. Deborah Wuttke). These research goals along with those of the minor users emphasizes the strength of biophysics and biochemistry at the University of Colorado, Boulder and the proposed instrument would fulfill a core need of a large community of users.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10RR026516-01
Application #
7792160
Study Section
Special Emphasis Panel (ZRG1-IMST-B (31))
Program Officer
Birken, Steven
Project Start
2010-07-01
Project End
2011-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
1
Fiscal Year
2010
Total Cost
$128,800
Indirect Cost
Name
University of Colorado at Boulder
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
007431505
City
Boulder
State
CO
Country
United States
Zip Code
80309
Ozdilek, Bagdeser A; Thompson, Valery F; Ahmed, Nasiha S et al. (2017) Intrinsically disordered RGG/RG domains mediate degenerate specificity in RNA binding. Nucleic Acids Res 45:7984-7996
Wostenberg, Christopher; Ceres, Pablo; Polaski, Jacob T et al. (2015) A Highly Coupled Network of Tertiary Interactions in the SAM-I Riboswitch and Their Role in Regulatory Tuning. J Mol Biol 427:3473-3490
Trausch, Jeremiah J; Batey, Robert T (2014) A disconnect between high-affinity binding and efficient regulation by antifolates and purines in the tetrahydrofolate riboswitch. Chem Biol 21:205-16
Stoddard, Colby D; Widmann, Jeremy; Trausch, Jeremiah J et al. (2013) Nucleotides adjacent to the ligand-binding pocket are linked to activity tuning in the purine riboswitch. J Mol Biol 425:1596-611
Garst, Andrew D; Porter, Ely B; Batey, Robert T (2012) Insights into the regulatory landscape of the lysine riboswitch. J Mol Biol 423:17-33
Fiegland, Larry R; Garst, Andrew D; Batey, Robert T et al. (2012) Single-molecule studies of the lysine riboswitch reveal effector-dependent conformational dynamics of the aptamer domain. Biochemistry 51:9223-33
Altschuler, Sarah E; Croy, Johnny E; Wuttke, Deborah S (2012) A small molecule inhibitor of Pot1 binding to telomeric DNA. Biochemistry 51:7833-45
Johnson Jr, James E; Reyes, Francis E; Polaski, Jacob T et al. (2012) B12 cofactors directly stabilize an mRNA regulatory switch. Nature 492:133-7
Trausch, Jeremiah J; Ceres, Pablo; Reyes, Francis E et al. (2011) The structure of a tetrahydrofolate-sensing riboswitch reveals two ligand binding sites in a single aptamer. Structure 19:1413-23