Abstract

We have developed an algorithm for structure-based prediction of the effects of temperature on electrostatic interactions in proteins. This algorithm has been adapted for calculation of the pH dependence of the transition temperature (Tm) of proteins. The approach uses the G determined at one pH value by DSC as a reference point. The pH-dependent component of the free energy of stabilization of the native state is calculated from structure and used to """"""""correct"""""""" the calorimetric G in order to account for the effects of pH on the stability of the protein. In order to test the algorithm, we have measured Tm vs pH for horse heart cytochrome-c over a range of conditions of ionic strength.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR004328-10
Application #
6122044
Study Section
Project Start
1997-08-05
Project End
1998-08-04
Budget Start
Budget End
Support Year
10
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Type
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Jaganaman, Sunil; Pinto, Alex; Tarasev, Michael et al. (2007) High levels of expression of the iron-sulfur proteins phthalate dioxygenase and phthalate dioxygenase reductase in Escherichia coli. Protein Expr Purif 52:273-9
Todd, M J; Gomez, J (2001) Enzyme kinetics determined using calorimetry: a general assay for enzyme activity? Anal Biochem 296:179-87
Karantza, V; Freire, E; Moudrianakis, E N (2001) Thermodynamic studies of the core histones: stability of the octamer subunits is not altered by removal of their terminal domains. Biochemistry 40:13114-23
Griko, Y V; Remeta, D P (1999) Energetics of solvent and ligand-induced conformational changes in alpha-lactalbumin. Protein Sci 8:554-61
Koder, R L; Miller, A F (1998) Overexpression, isotopic labeling, and spectral characterization of Enterobacter cloacae nitroreductase. Protein Expr Purif 13:53-60
Freire, E (1998) Statistical thermodynamic linkage between conformational and binding equilibria. Adv Protein Chem 51:255-79
Lee, R T; Gabius, H J; Lee, Y C (1998) Thermodynamic parameters of the interaction of Urtica dioica agglutinin with N-acetylglucosamine and its oligomers. Glycoconj J 15:649-55
Zakharov, S D; Lindeberg, M; Griko, Y et al. (1998) Membrane-bound state of the colicin E1 channel domain as an extended two-dimensional helical array. Proc Natl Acad Sci U S A 95:4282-7
Luque, I; Todd, M J; Gomez, J et al. (1998) Molecular basis of resistance to HIV-1 protease inhibition: a plausible hypothesis. Biochemistry 37:5791-7
Chu, V; Freitag, S; Le Trong, I et al. (1998) Thermodynamic and structural consequences of flexible loop deletion by circular permutation in the streptavidin-biotin system. Protein Sci 7:848-59

Showing the most recent 10 out of 86 publications