[12] Fluorescence anisotropy applied to biomolecular interactions

Publisher Summary This chapter focuses on the applications of polarization or anisotropy data. The appeal of fluorescence spectroscopy in the study of biomolecular systems lies in the characteristic time scale of the emission process, the sensitivity of the technique, and its ability to accommodate rapid and facile changes in the solvent milieu under conditions corresponding to thermodynamic equilibrium. On excitation of a fluorescent solution and observation of the emission at right angles both to the direction of propagation of the exciting light and to the direction of the electric vector, the polarization of the emission is defined in this chapter. The optical densities of the solution may be unavoidably high (even allowing for smaller cuvette or front-face geometries), and the requisite inner filter corrections may lower confidence in the binding data. Polarization/anisotropy measurements—such as lifetime measurements—are intensive quantities and as such are not subject to first-order inner filter corrections.

[1]  K. Rajkowski,et al.  Corrected equations for the calculation of protein-ligand binding results from fluorescence polarization data. , 1981, Journal of theoretical biology.

[2]  K. Sakumi,et al.  Interaction of Ada protein with DNA examined by fluorescence anisotropy of the protein. , 1990, Biochemistry.

[3]  R. Steiner,et al.  Excited states of proteins and nucleic acids , 1972 .

[4]  T. Dewey,et al.  Biophysical and Biochemical Aspects of Fluorescence Spectroscopy , 2013, Springer US.

[5]  M. Chabbert,et al.  Kinetic studies of recA protein binding to a fluorescent single-stranded polynucleotide. , 1987, Biochemistry.

[6]  D. Rousseau.  Optical techniques in biological research , 1984 .

[7]  J. Haralambidis,et al.  The synthesis of polyamide-oligonucleotide conjugate molecules. , 1990, Nucleic acids research.

[8]  Edward W. Voss,et al.  Fluorescein hapten : an immunological probe , 1984 .

[9]  M. Shinitzky,et al.  Fluorescence polarization studies of lysozyme and lysozyme-saccharide complexes. , 1973, Journal of molecular biology.

[10]  J. Jentoft,et al.  Interactions at the nucleic acid binding site of the avian retroviral nucleocapsid protein: studies utilizing the fluorescent probe 4,4'-bis(phenylamino)(1,1'-binaphthalene)-5,5'-disulfonic acid. , 1990, Biochemistry.

[11]  D. Laurence,et al.  A study of the adsorption of dyes on bovine serum albumin by the method of polarization of fluorescence. , 1952, The Biochemical journal.

[12]  J. Haralambidis,et al.  The preparation of polyamide-oligonucleotide probes containing multiple non-radioactive labels. , 1990, Nucleic acids research.

[13]  C. Royer,et al.  Role of protein--protein interactions in the regulation of transcription by trp repressor investigated by fluorescence spectroscopy. , 1992, Biochemistry.

[14]  J. Beechem,et al.  Resolution of the fluorescence decay of the two tryptophan residues of lac repressor using single tryptophan mutants. , 1990, Biophysical journal.

[15]  J. Villafranca,et al.  Interactive binding between the substrate and allosteric sites of carbamoyl-phosphate synthetase. , 1988, Biochemistry.

[16]  C. A. Parker.  Fluorescence and Phosphorescence Analysis , 1967 .

[17]  S. Benkovic,et al.  Fluorescent oligonucleotides and deoxynucleotide triphosphates: preparation and their interaction with the large (Klenow) fragment of Escherichia coli DNA polymerase I. , 1989, Biochemistry.

[18]  D. Malencik,et al.  High affinity binding of the mastoparans by calmodulin. , 1983, Biochemical and biophysical research communications.

[19]  E. Gratton,et al.  Intrinsic fluorescence of elongation factor Tu in its complexes with GDP and elongation factor Ts. , 1987, Biochemistry.

[20]  G. Weber.  Polarization of the fluorescence of macromolecules. I. Theory and experimental method. , 1952, The Biochemical journal.

[21]  D. Malencik,et al.  Association of melittin with the isolated myosin light chains. , 1988, Biochemistry.

[22]  C. Royer,et al.  Solution studies of the interactions between the histone core proteins and DNA using fluorescence spectroscope , 1992 .

[23]  P. Bally,et al.  Estimation of the saturation fraction of bound fluorescent ligands from the intensity and anisotropy of the emission , 1980 .

[24]  F. Perrin.  Polarisation de la lumière de fluorescence. Vie moyenne des molécules dans l'etat excité , 1926 .

[25]  S. Benkovic,et al.  Interaction of DNA with the Klenow fragment of DNA polymerase I studied by time-resolved fluorescence spectroscopy. , 1991, Biochemistry.

[26]  S. Benkovic,et al.  Resonance energy transfer measurements between substrate binding sites within the large (Klenow) fragment of Escherichia coli DNA polymerase I. , 1989, Biochemistry.

[27]  S. Benkovic,et al.  Synthesis and application of derivatizable oligonucleotides. , 1987, Nucleic acids research.

[28]  A. Murakami,et al.  Fluorescent-labeled oligonucleotide probes: detection of hybrid formation in solution by fluorescence polarization spectroscopy. , 1991, Nucleic acids research.

[29]  G W Tregear,et al.  Preparation of base-modified nucleosides suitable for non-radioactive label attachment and their incorporation into synthetic oligodeoxyribonucleotides. , 1987, Nucleic acids research.

[30]  E. Geiduschek,et al.  A type II DNA-binding protein genetically engineered for fluorescence spectroscopy: the "arm" of transcription factor 1 binds in the DNA grooves. , 1989, Biochemistry.

[31]  D. Millar,et al.  Dynamics of mismatched base pairs in DNA. , 1991, Biochemistry.

[32]  W. Dandliker,et al.  Quantification of the antigen-antibody reaction by the polarization of fluorescence. , 1961, Biochemical and biophysical research communications.

[33]  A fluorescence study of the binding of eucaryotic initiation factors to messenger RNA and messenger RNA analogues. , 1987, Biochemistry.