The long-term objective of the proposed research is the elucidation of the effects of pressure on the dynamics and structure of proteins and biological membranes. Pressure is a fundamental thermodynamic variable that provides information on processes that involve changes in volume, including the folding and unfolding of proteins, lipid phase changes in membranes, and the modulation of the fluidity and interfacial properties of membranes. The proposed work focuses on the effects of pressure on the dynamics, phase state, and conformation of several model systems: homopolypeptides (poly-L-lysine, poly-L-glutamic acid, and poly-N -(3- hydroxypropyl)-L-glutamine), hen egg-white lysozyme, sperm whale myoglobin, and various pure phospholipids in the form of sonicated vesicles or multilamellar dispersions. A variety of advanced high resolution NMR methods, using H, H, P nuclei, will be employed together with unique high-pressure NMR probes, in the range from 1 bar to 1O kbar and from 0 to, 100 degree C. In the proteins and the homopolypeptides, selected 1H and 13C resonances will be used to study the dynamics in various regions of the macromolecules, and to examine the thermodynamics and kinetics of the folding-unfolding transitions. The dynamics of phospholipid molecules in liposomes will be studied by 13C NMR: the order and con-formation of the acyl chains and the head groups will be investigated by 13C-NMR, using deuterated lipids, and by H-2D NMR techniques: and phase changes will be examined by 31P and H NMR methods. In addition, slow motions and the lateral diffusion of phospholipids in liposomes will be investigated by rotating frame NMR relaxation techniques.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042452-04
Application #
3301010
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1989-07-01
Project End
1999-06-30
Budget Start
1992-07-01
Budget End
1993-06-30
Support Year
4
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
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

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