Goal of the proposal is the detection and characterization of segmental motions, structural domains and solvent effects on the shape, dimensions, polymerization, and biological functions of soluble and membrane-bound proteins. Segmental motions will be monitored using fluorescence depolarization techniques. The shape and dimension of protein molecules will be inferred from their rotational diffusion and from measurements of energy transfer of fluorescence. The presence of structural domains will be investigated using controlled unfolding processes in denaturing agent-like guanidine hydrochloride, pH and temperature. Removal of prosthetic groups, like heme from hemoglobin, also produce partial unfoldings, which may reveal the presence of structural domains. The proteins to be investigated at the start of the project are hemoglobin and its heme-free derivative, band 3 of the erythrocyte ghosts and calcium dependent ATPase of skeletal sarcoplasmic reticulum. The investigation will increase our understanding of the mechanism that requlate biological transport of gases and ions in fluids and across membranes.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL013164-15
Application #
3334587
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1977-04-01
Project End
1988-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
15
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Maryland Baltimore
Department
Type
Schools of Medicine
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201
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Fronticelli, C; Brinigar, W S; Olson, J S et al. (1993) Recombinant human hemoglobin: modification of the polarity of the beta-heme pocket by a valine67(E11)-->threonine mutation. Biochemistry 32:1235-42
Gryczynski, Z; Fronticelli, C; Tenenholz, T et al. (1993) Effect of disordered hemes on energy transfer rates between tryptophans and heme in myoglobin. Biophys J 65:1951-8
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