The general aim in this project is to apply physicochemical techniques and principles to a wide variety of biochemical and biological processes. Although our efforts will continue, as in the past, to be centered on the determination and interpretation of the thermodynamics of such processes, it is frequently found that significant non-thermodynamic results are obtained. Our research may be illustrated by the following examples. (a) Differential scanning calorimetry (DSC) affords a powerful tool for the study of the interactions of a broad range of substance with lipids in bilayer suspension serving as models for biological membranes. We are using DSC to investigate the effects on lipid bilayers of small molecules including drugs of various types, and of proteins. (b) Recent developments indicate that careful DSC studies of protein denaturation can give important information concerning the structures of proteins, in particular the division of these structures into more or less independent domains. (c) Isothermal calorimetry provides the best method for the determination of the enthalpy and heat capacity changes in biochemical processes. We have recently used this technique in studies of the binding of FMN to phosphorylase a and of glucose to hexokinase. (d) We have demonstrated that DSC is potentially very useful in the study of cellular processes such as phagocytosis, not only for the evaluation of the energetics of the process but also in obtaining new insights into the mechanisms involved.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM004725-31
Application #
3267854
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1974-10-15
Project End
1988-02-29
Budget Start
1986-12-01
Budget End
1988-02-29
Support Year
31
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Yale University
Department
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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