The long-term objective of the proposed research is to contribute to the understanding of the stability, folding intermediates, and dynamics of proteins by using pressure as the fundamental thermodynamic variable. Unique high pressure equipment, suitable for measurements from 0.1 MPa (l bar) to l GPa (10 kbar) and from -25 degrees C to lOO degrees C, will be used in conjunction with a variety of l-D and 2-D high resolution NMR techniques to study the effects of pressure on the unfolding/folding equilibria, conformational states of native and compact denatured states, and the dynamic properties of several model proteins. In addition, the cold-denaturation behavior of these proteins will be studied by taking advantage of the depression of the freezing point of water by pressure. The pressure, cold, heat, and chemically denatured states of the model proteins will be examined and compared, with special emphasis on the compact denatured states observed by cold and pressure denaturation. The model proteins selected for these studies include hen egg-white lysozyme, bacteriophage T4 lysozyme, horse myoglobin, sperm-whale myoglobin, staphylococcal nuclease, and bovine pancreatic ribonuclease A, as well as selected mutants of T4 lysozyme, sperm-whale myoglobin, staphylococcal nuclease, and bovine pancreatic ribonuclease A. To complement the NMR measurements at high pressure, especially on the compact denatured states of the model proteins and their mutants, circular dichroism (CD) and laser-photo chemically induced dynamic nuclear polarization (CIDNP) experiments will be performed. High-pressure CD cells and CIDNP probes will be designed and constructed.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM042452-06
Application #
2181382
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1989-07-01
Project End
1998-07-31
Budget Start
1994-08-01
Budget End
1995-07-31
Support Year
6
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Bondos, S E; Sligar, S; Jonas, J (2000) High-pressure denaturation of apomyoglobin. Biochim Biophys Acta 1480:353-64
Yu, A; Ballard, L; Smillie, L et al. (1999) Effects of high pressure and temperature on the wild-type and F29W mutant forms of the N-domain of avian troponin C. Biochim Biophys Acta 1431:53-63
Jonas, J; Ballard, L; Nash, D (1998) High-resolution, high-pressure NMR studies of proteins. Biophys J 75:445-52
Ballard, L; Yu, A; Reiner, C et al. (1998) A high-pressure, high-resolution NMR probe for experiments at 500 MHz. J Magn Reson 133:190-3
Nash, D P; Jonas, J (1997) Structure of pressure-assisted cold denatured lysozyme and comparison with lysozyme folding intermediates. Biochemistry 36:14375-83
Nash, D P; Jonas, J (1997) Structure of the pressure-assisted cold denatured state of ubiquitin. Biochem Biophys Res Commun 238:289-91
Nash, D; Lee, B S; Jonas, J (1996) Hydrogen-exchange kinetics in the cold denatured state of ribonuclease A. Biochim Biophys Acta 1297:40-8
Peng, X; Jonas, A; Jonas, J (1995) High pressure 2H-NMR study of the order and dynamics of selectively deuterated dipalmitoyl phosphatidylcholine in multilamellar aqueous dispersions. Biophys J 68:1137-44
Zhang, J; Peng, X; Jonas, A et al. (1995) NMR study of the cold, heat, and pressure unfolding of ribonuclease A. Biochemistry 34:8631-41
Peng, X; Jonas, A; Jonas, J (1995) One and two dimensional 1H-NMR studies of pressure and tetracaine effects on sonicated phospholipid vesicles. Chem Phys Lipids 75:59-69

Showing the most recent 10 out of 18 publications