Abstract

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.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL013164-16
Application #
3334588
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1977-04-01
Project End
1989-03-31
Budget Start
1987-12-01
Budget End
1989-03-31
Support Year
16
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
University of Maryland Baltimore
Department
Type
Schools of Medicine
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201

  Publications

Thomas, M; Matheson-Urbaitis, B; Kwansa, H et al. (1997) Introduction of negative charges to a crosslinked hemoglobin: lack of effect on plasma half time. Artif Cells Blood Substit Immobil Biotechnol 25:309-14
Bucci, E; Razynska, A; Kwansa, H et al. (1996) Production and characteristics of an infusible oxygen-carrying fluid based on hemoglobin intramolecularly cross-linked with sebacic acid. J Lab Clin Med 128:146-53
Bucci, E; Razynska, A; Kwansa, H et al. (1996) Positive and negative cooperativities at subsequent steps of oxygenation regulate the allosteric behavior of multistate sebacylhemoglobin. Biochemistry 35:3418-25
Razynska, A; Matheson-Urbaitis, B; Fronticelli, C et al. (1996) Stabilization of the tetrameric structure of human and bovine hemoglobins by pseudocrosslinking with muconic acid. Arch Biochem Biophys 326:119-25
Matheson-Urbaitis, B; Lu, Y S; Fronticelli, C et al. (1995) Renal and systemic-hemodynamic response to isovolemic exchange transfusion with hemoglobin cross-linked with bis (3,5-dibromosalicyl) fumarate or albumin. J Lab Clin Med 126:250-60
Fronticelli, C; Gattoni, M; Lu, A L et al. (1994) The dimer-tetramer equilibrium of recombinant hemoglobins. Stabilization of the alpha 1 beta 2 interface by the mutation beta(Cys112-->Gly) at the alpha 1 beta 1 interface. Biophys Chem 51:53-7
Fronticelli, C; Pechik, I; Brinigar, W S et al. (1994) Chloride ion independence of the Bohr effect in a mutant human hemoglobin beta (V1M+H2deleted). J Biol Chem 269:23965-9
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
Gryczynski, Z; Bucci, E; Kuyyba, J (1993) Linear dichroism study of metalloporphyrin transition moments in view of radiationless interactions with tryptophan in hemoproteins. Photochem Photobiol 58:492-8

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