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Abstract:

This work describes a laboratory experiment to illustrate the usefulness of fluorescence anisotropy in the field of biophysics. Fluorescence anisotropy of dansyl-labeled BSA was determined in media of increasing glycerol concentrations. The Perrin equation was fitted to the experimental data, obtaining the molecular volume of the protein. The simplicity of the experiment and data analysis helped the students to focus on the relationship between probe anisotropy and rotational diffusion. Additionally, this laboratory exercise has the advantage of using a protein and a probe that are inexpensive and very common in many laboratories. © 2003 by The International Union of Biochemistry and Molecular Biology.

Registro:

Documento: Artículo
Título:Determination of the molecular size of BSA by fluorescence anisotropy
Autor:Flecha, F.L.G.; Levi, V.
Filiación:Instituto de Química y Fisicoquímica Biológicas, Departamento de Química Biológica, Universidad de Buenos Aires, Argentina
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, United States
Palabras clave:BSA; Fluorescence anisotropy; Molecular size; glycerol; protein; protein BSA; unclassified drug; anisotropy; article; biophysics; data analysis; fluorescence analysis; molecular probe; molecular size; university student
Año:2003
Volumen:31
Número:5
Página de inicio:319
Página de fin:322
DOI: http://dx.doi.org/10.1002/bmb.2003.494031050261
Título revista:Biochemistry and Molecular Biology Education
Título revista abreviado:Biochem. Mol. Biol. Educ.
ISSN:14708175
CODEN:BMBEC
CAS:glycerol, 56-81-5; protein, 67254-75-5
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14708175_v31_n5_p319_Flecha

Referencias:

  • Feynman, R.P., Leighton, R.B., Sands, M.L., (1963) The Feymann Lectures on Physics, Mainly Mechanics, Radiation and Heat, p. 33. , Addison-Wesley, Reading, Massachusetts
  • Cantor, C.R., Schimmel, P.R., (1980) Biophysical Chemistry, pp. 125-167. , W. H. Freeman, New York
  • Valeur, B., (2001) Molecular Fluorescence: Principles and Applications, pp. 125-167. , Wiley, New York
  • Weber, G., Rotational Brownian motion and polarization of the fluorescence of solutions (1953) Adv. Protein Chem., 8, pp. 415-459
  • Van Holde, K.E., Johnson, W.C., Ho, P.S., (1998) Principles of Physical Biochemistry, pp. 452-483. , Prentice Hall, New Jersey
  • Weber, G., Polarization of the fluorescence of macromolecules. I. Theory and experimental method (1952) Biochem. J., 51, pp. 145-155
  • Baud, S., Margeat, E., Lumbroso, S., Paris, F., Sultan, C., Royer, C., Poujol, N., Equilibrium binding assays reveal the elevated stoichiometry and salt dependence of the interaction between full-length human sex-determining region on the Y chromosome (SRY) and DNA (2002) J. Biol. Chem., 277, pp. 18404-18410
  • Engel, M.F., Van Mierlo, C.P., Visser, A.J., Kinetic and structural characterization of adsorption-induced unfolding of bovine alpha-lactalbumin (2002) J. Biol. Chem., 277, pp. 10922-10930
  • Levi, V., Gonzalez Flecha, F.L., Labeling of proteins with fluorescent probes. Photophysical characterization of dansylated bovine serum albumin (2003) Biochem. Mol. Biol. Educ., 31, pp. 333-336
  • Seber, G.A.F., Wild, C.J., (1989) Nonlinear Regression, pp. 21-27. , John Wiley & Sons, New York
  • Weber, G., Polarization of the fluorescence of macromolecules. II. Fluorescent conjugates of ovalbumin and bovine serum albumin (1952) Biochem. J., 51, pp. 155-167
  • Tanford, C., (1961) Physical Chemistry of Macromolecules, pp. 317-456. , Wiley, New York
  • Kuntz, I.D., Kauzmann, W., Hydration of proteins and polypeptides (1974) Adv. Protein Chem., 28, pp. 239-345
  • Axelsson, I., Characterization of proteins and other macromolecules by agarose gel chromatography (1978) J. Chromatogr. A, 152, pp. 21-32
  • Tinoco, I., Sauer, K., Wang, J.C., Puglisi, J.D., (2002) Physical Chemistry: Principles and Applications in Biological Sciences, , Prentice Hall, New Jersey
  • Levi, V., González Flecha, F.L., Reversible fast-dimerization of bovine serum albumin detected by fluorescence resonance energy transfer (2002) Biochim. Biophys. Acta., 1599, pp. 141-148
  • Jameson, D.M., Croney, J.C., Fluorescence polarization: Past, present, and future (2003) Comb-Chem-High Throughput Screen, 6, pp. 167-173

Citas:

---------- APA ----------
Flecha, F.L.G. & Levi, V. (2003) . Determination of the molecular size of BSA by fluorescence anisotropy. Biochemistry and Molecular Biology Education, 31(5), 319-322.
http://dx.doi.org/10.1002/bmb.2003.494031050261
---------- CHICAGO ----------
Flecha, F.L.G., Levi, V. "Determination of the molecular size of BSA by fluorescence anisotropy" . Biochemistry and Molecular Biology Education 31, no. 5 (2003) : 319-322.
http://dx.doi.org/10.1002/bmb.2003.494031050261
---------- MLA ----------
Flecha, F.L.G., Levi, V. "Determination of the molecular size of BSA by fluorescence anisotropy" . Biochemistry and Molecular Biology Education, vol. 31, no. 5, 2003, pp. 319-322.
http://dx.doi.org/10.1002/bmb.2003.494031050261
---------- VANCOUVER ----------
Flecha, F.L.G., Levi, V. Determination of the molecular size of BSA by fluorescence anisotropy. Biochem. Mol. Biol. Educ. 2003;31(5):319-322.
http://dx.doi.org/10.1002/bmb.2003.494031050261