The goal of this project is to investigate experimentally the physical basis of the stability of folded, native conformation of proteins and obtain quantitative information on the energetics of interactions determining this conformation. The lack of this information appears to be the main obstacle in understanding the principles of protein architecture and functioning, and, consequently, in designing the new proteins with the required structure and stability. The energetics of interactions in protein will be estimated by microcalorimetric studies of the processes of (a) unfolding/refolding of proteins with known three-dimensional structure and their mutant forms, and (b) transferring into water the low molecular substances modelling various groups in proteins. These experiments will be carried out in a broad temperature range and the use of new, precise reaction and scanning microcalorimetric techniques will enable us to determine the thermodynamic functions specifying these processes with much higher accuracy and completeness than has been possible before. Analyzing these functions, and especially their temperature dependencies, on the base of structural information on the folded and unfolded conformations, we can evaluate the contribution of hydration effects and intramolecular bonding in stabilization of the native protein structure and clarify, particularly, the mechanism of hydrophobic interactions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM048036-02
Application #
3307470
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1992-08-01
Project End
1996-07-31
Budget Start
1993-08-01
Budget End
1994-07-31
Support Year
2
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Arts and Sciences
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Privalov, Peter L; Dragan, Anatoly I; Crane-Robinson, Colyn (2009) The cost of DNA bending. Trends Biochem Sci 34:464-70
Crane-Robinson, Colyn; Dragan, Anatoly I; Read, Christopher M (2009) Defining the thermodynamics of protein/DNA complexes and their components using micro-calorimetry. Methods Mol Biol 543:625-51
Dragan, Anatoly I; Privalov, Peter L (2008) Use of fluorescence resonance energy transfer (FRET) in studying protein-induced DNA bending. Methods Enzymol 450:185-99
Privalov, Peter L; Dragan, Anatoly I (2007) Microcalorimetry of biological macromolecules. Biophys Chem 126:16-24
Privalov, Peter L; Dragan, Anatoly I; Crane-Robinson, Colyn et al. (2007) What drives proteins into the major or minor grooves of DNA? J Mol Biol 365:1-9
Crane-Robinson, Colyn; Dragan, Anatoly I; Privalov, Peter L (2006) The extended arms of DNA-binding domains: a tale of tails. Trends Biochem Sci 31:547-52
Dragan, Anatoly I; Li, Zhenlan; Makeyeva, Elena N et al. (2006) Forces driving the binding of homeodomains to DNA. Biochemistry 45:141-51
Hargreaves, Victoria V; Makeyeva, Elena N; Dragan, Anatoly I et al. (2005) Stability and DNA binding ability of the DNA binding domains of interferon regulatory factors 1 and 3. Biochemistry 44:14202-9
Dragan, Anatoly I; Read, Christopher M; Makeyeva, Elena N et al. (2004) DNA binding and bending by HMG boxes: energetic determinants of specificity. J Mol Biol 343:371-93
Dragan, Anatoly I; Potekhin, Sergey A; Sivolob, Andrei et al. (2004) Kinetics and thermodynamics of the unfolding and refolding of the three-stranded alpha-helical coiled coil, Lpp-56. Biochemistry 43:14891-900

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