We will attempt to gain information about the motional behavior of proteins by studying the ability of these motions to allow a variety of small molecules to come into contact with normally buried protein groups. Target positions within the proteins will be fluorescent and phosphorescent tryptophans, porphyrins, and individual identifiable protons. Collisional encounters will be registered by the quenching of fluorescence and phosphorescence, by phosphorescence detected H-D exchange, and by 1H NMR T1 measurements. A wide range of proteins and small molecule agents will be studied. These methods are expected to distinguish between and partially characterize: 1) the small rapid liquid-like motions that may create transient channels accommodating the entry into the protein matrix of the small test molecules; 2) more sizeable opening reactions of the kind that may bring buried groups out into contact with the solvent. In addition other factors that may operate in governing protein-small molecule interactions will be studied, including the possibility of binding of the small molecules to and diffusion over the protein surface and the role of partial exposure rather than full burial.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM034448-04
Application #
3285465
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1984-12-01
Project End
1991-01-31
Budget Start
1988-02-01
Budget End
1989-01-31
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Laberge, M; Wright, W W; Sudhakar, K et al. (1997) Conformational effects of calcium release from parvalbumin: comparison of computational simulations with spectroscopic investigations. Biochemistry 36:5363-71
Angiolillo, P J; Vanderkooi, J M (1996) Hydrogen atoms are produced when tryptophan within a protein is irradiated with ultraviolet light. Photochem Photobiol 64:492-5
Jez, J M; Vanderkooi, J M; Laties, A M (1996) Spectroscopic characterization of bendazac and benzydamine: possible photochemical modes of action. Biochem Biophys Res Commun 221:266-70
Sudhakar, K; Erecinska, M; Vanderkooi, J M (1995) Interaction of polyamines with the Ca(2+)-binding protein parvalbumin. Eur J Biochem 230:498-502
Angiolillo, P J; Vanderkooi, J M (1995) Electron paramagnetic resonance of the excited triplet state of metal-free and metal-substituted cytochrome c. Biophys J 68:2505-18
Sudhakar, K; Phillips, C M; Owen, C S et al. (1995) Dynamics of parvalbumin studied by fluorescence emission and triplet absorption spectroscopy of tryptophan. Biochemistry 34:1355-63
Sudhakar, K; Phillips, C M; Williams, S A et al. (1993) Excited states of tryptophan in cod parvalbumin. Identification of a short-lived emitting triplet state at room temperature. Biophys J 64:1503-11
Wright, W W; Owen, C S; Vanderkooi, J M (1992) Penetration of analogues of H2O and CO2 in proteins studied by room temperature phosphorescence of tryptophan. Biochemistry 31:6538-44
Dadak, V; Vanderkooi, J M; Wright, W W (1992) Electron transfer from excited tryptophan to cytochrome c: mechanism of phosphorescence quenching? Biochim Biophys Acta 1100:33-9
Papp, S; Vanderkooi, J M; Owen, C S et al. (1990) Reactions of excited triplet states of metal substituted myoglobin with dioxygen and quinone. Biophys J 58:177-86

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