Abstract

Steady-state and nanosecond time-resolved fluorescence methods will be used to investigate the kinetics of excited-state processes of interest in biology. Fluorescence decay, time-resolved emission spectra and time-resolved fluorescence emission anisotropy will be used to study solvent relaxation, proton transfer, energy transfer and rotational motion of fluorophores. Instrumental methods for obtaining the required data and computational procedures for data analysis will be improved. Fluorophores will be studied in solution and when bound to proteins and membrane systems. The fluorescence of tryptophan peptides and tryptophan in proteins will be investigated. Fluorescence decay will be used to uncover microheterogeneity. Information will also be obtained about excited state interactions between tryptophan and solvent molecules. Decay of the fluorescence emission anisotropy will be used to study the rotational behavior of proteins and peptides as well as segmental motion of tryptophan residues. A general aim of the studies is to discover in what way dynamic interactions on the nanosecond time scale are related to the functions of membranes and proteins.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM011632-23
Application #
3268280
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1977-09-01
Project End
1987-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
23
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Type
Schools of Arts and Sciences
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Brown, M P; Shaikh, N; Brenowitz, M et al. (1994) The allosteric interaction between D-galactose and the Escherichia coli galactose repressor protein. J Biol Chem 269:12600-5
Wu, P; Li, Y K; Talalay, P et al. (1994) Characterization of the three tyrosine residues of delta 5-3-ketosteroid isomerase by time-resolved fluorescence and circular dichroism. Biochemistry 33:7415-22
Wu, P; Brand, L (1994) Resonance energy transfer: methods and applications. Anal Biochem 218:1-13
Chauvin, F; Brand, L; Roseman, S (1994) Sugar transport by the bacterial phosphotransferase system. Characterization of the Escherichia coli enzyme I monomer/dimer transition kinetics by fluorescence anisotropy. J Biol Chem 269:20270-4
Wu, P; Brand, L (1994) Conformational flexibility in a staphylococcal nuclease mutant K45C from time-resolved resonance energy transfer measurements. Biochemistry 33:10457-62
Chauvin, F; Brand, L; Roseman, S (1994) Sugar transport by the bacterial phosphotransferase system. Characterization of the Escherichia coli enzyme I monomer/dimer equilibrium by fluorescence anisotropy. J Biol Chem 269:20263-9
Hirshfield, K M; Toptygin, D; Packard, B S et al. (1993) Dynamic fluorescence measurements of two-state systems: applications to calcium-chelating probes. Anal Biochem 209:209-18
Wu, P G; James, E; Brand, L (1993) Compact thermally-denatured state of a staphylococcal nuclease mutant from resonance energy transfer measurements. Biophys Chem 48:123-33
Toptygin, D; Brand, L (1993) Fluorescence decay of DPH in lipid membranes: influence of the external refractive index. Biophys Chem 48:205-20
Rice, K G; Wu, P; Brand, L et al. (1993) Modification of oligosaccharide antenna flexibility induced by exoglycosidase trimming. Biochemistry 32:7264-70

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