The broad aim in this project is the study of biological and biochemical processes using physicochemical techniques. As in the past, our efforts will be centered on the determination, primarily by calorimetric methods and the interpretation of the thermodynamics of such processes. A major fraction of our work will involve study of thermally-induced processes in proteins, nucleic acids and lipids by means of high sensitivity differential scanning calorimetry (DSC). The power and applicability of this technique is at present rapidly expanding. For example, DSC affords the best means for checking the thermal stability of proteins engineered to perform some specific pharmaceutical function, and for acquiring thermodynamic data which may suggest specific ways to improve the thermal stability. Our DSC work on proteins will remain centered on studying the effects of mutations using materials supplied to use by a wide range of collaborators. Included will be, among others, staphylococcal nuclease, streptomyces subtilisin inhibitor (SSI), thioredoxin and cytochrome b562. DSC work on nucleotides will include study of the unfolding of synthetic triple stranded DNA, and of various nucleotide-protein complexes also studies by ITC. DSC gives an excellent means for studying the phase transitions of lipids. Work on pure synthetic lipids of unusual structure will continue. Studies of the effects of added materials (Ca++, Mg++, AlCl3, nitrophenyl esters) on the thermotropic behavior of lipid bilayers will also be continued. We are making constantly increasing use of isothermal situation calorimetry (ITC), in studying the binding of ligands to proteins and nucleic acids, the interactions of proteins with nucleic acids, for example, the trp repressor-trp operator interaction, and other biochemical reactions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM004725-40
Application #
2166059
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1974-10-15
Project End
1998-11-30
Budget Start
1995-12-01
Budget End
1998-11-30
Support Year
40
Fiscal Year
1996
Total Cost
Indirect Cost
Name
Yale University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Thomson, J; Liu, Y; Sturtevant, J M et al. (1998) A thermodynamic study of the binding of linear and cyclic oligosaccharides to the maltodextrin-binding protein of Escherichia coli. Biophys Chem 70:101-8
Robinson, C R; Liu, Y; O'Brien, R et al. (1998) A differential scanning calorimetric study of the thermal unfolding of apo- and holo-cytochrome b562. Protein Sci 7:961-5
Robinson, C R; Liu, Y; Thomson, J A et al. (1997) Energetics of heme binding to native and denatured states of cytochrome b562. Biochemistry 36:16141-6
Lemmon, M A; Bu, Z; Ladbury, J E et al. (1997) Two EGF molecules contribute additively to stabilization of the EGFR dimer. EMBO J 16:281-94
Liu, Y; Sturtevant, J M (1996) The observed change in heat capacity accompanying the thermal unfolding of proteins depends on the composition of the solution and on the method employed to change the temperature of unfolding. Biochemistry 35:3059-62
DeDecker, B S; O'Brien, R; Fleming, P J et al. (1996) The crystal structure of a hyperthermophilic archaeal TATA-box binding protein. J Mol Biol 264:1072-84
Mandiyan, V; O'Brien, R; Zhou, M et al. (1996) Thermodynamic studies of SHC phosphotyrosine interaction domain recognition of the NPXpY motif. J Biol Chem 271:4770-5
Tanaka, A (1996) Steady-state kinetic and calorimetric studies on the binding of Aspergillus niger glucoamylase with gluconolactone, 1-deoxynojirimycin, and beta-cyclodextrin. Biosci Biotechnol Biochem 60:2055-8
O'Brien, R; Sturtevant, J M; Wrabl, J et al. (1996) A scanning calorimetric study of unfolding equilibria in homodimeric chicken gizzard tropomyosins. Biophys J 70:2403-7
Tamura, A; Sturtevant, J M (1995) A thermodynamic study of mutant forms of Streptomyces subtilisin inhibitor. I. Hydrophobic replacements at the position of Met103. J Mol Biol 249:625-35

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