Abstract
Circular dichroism (CD) is the difference in absorption of left and right circularly polarized light, usually by a solution containing the molecules of interest. A signal is only measured for chiral molecules such as proteins. A CD spectrum provides information about the bonds and structures responsible for this chirality. When a small molecule (or ligand) binds to a protein, it acquires an induced CD (ICD) spectrum through chiral perturbation to its structure or electron rearrangements. The wavelengths of this ICD are determined by the ligand’s own absorption spectrum, and the intensity of the ICD spectrum is determined by the strength and geometry of its interaction with the protein. Thus, ICD can be used to probe the binding of ligands to proteins. This chapter outlines protein CD and ICD, together with some of the issues relating to experimental design and implementation.
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References
Nakanishi K., Berova N., and Woody R. W. (ed.) (1994) Circular Dichroism: Principles and Applications. VCH, New York.
Rodger A. and Nordén B (1997) Circular and Linear Dichroism. Oxford University Press, Oxford.
Johnson W. C. Fitting programs are available to deconvolute the experimental data into percentages of the structural motifs. Website: e.g., http://www.cryst. bbk.ac.uk/cdweb/html/home.html; Dicroweb: a facility of the BBSRC Centre for Protein and Membrane Structure and Dynamics. http://oregonstate.edu/dept/ biochem/faculty/johnson.html. Date accessed: November 20, 2004.
Johnson W. C. (1999) Analyzing protein circular dichroism spectra for accurate secondary structures. Proteins: Structure, Function and Genetics 7, 307–312.
Chen G. C. and Yang J. T. (1977) Two-point calibration of circular dichrometer with D-10-camphorsulfonic acid. Anal. Lett. 10, 1195–1207.
Takakuwa T., Konno T., and Meguro H. (1985) A new standard substance for calibration of circular dichroism: ammonium D-10-camphorsulfonate. Anal. Sci. 1, 215–218.
Dafforn T. R., Halsall D. J., Serpell L. C., Rajendra J., and Rodger A. (2004) The use of linear dichroism to determine the orientation of secondary structural elements within protein fibres. Biophys. J. 86, 404–410.
Pace C. N. (1986) Determination and analysis of urea and guanidine hydrochloride denaturation curves Methods Enzymol. 131, 266–280.
Gill S. C. and von Hippel P. H. (1989) Calculation of protein extinction coefficients from amino acid sequence data. Anal. Biochem. 182, 319–326.
Miles A. J., Wien F., Lees J. G., Rodger A., Janes R. W., and Wallace B. A. (2003) Calibration and standardisation of synchrotron radiation circular dichroism and conventional circular dichroism spectrophotometers. Spectroscopy 17, 653–661.
Scatchard G. (1949) The attraction of proteins for small molecules and ions. Ann. N.Y. Acad. Sci. 51, 660–672.
Rodger A. (1993) Linear dichroism. Methods Enzymol. 226, 232–258.
Polster J. and Lachman H. (1989) Spectrometric Titrations: Analysis of Chemical Equilibria. VCH Verlagsgesellschaft, Weinheim, Germany.
Johnson W. C., Jr. (1988) Secondary structure of proteins through circular dichroism spectroscopy. Ann. Rev. Biophys. Biophys. Chem. 17, 145–166.
Johnson W. C., Jr. (1985) Circular dichroism and its empirical application to biopolymers. Methods Biochem. Anal. 31, 61–163.
Miguel M. S., Marrington R., Rodger P. M., Rodger A., and Robinson C. (2003) An Escherichia coli twin-arginine signal peptide switches between helical and unstructured conformations depending on hydrophobicity of the environment. Euro. J. Biochem. 270, 1–8.
Green P. (1999) PhD Thesis, University of Warwick, UK.
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© 2005 Humana Press Inc., Totowa, NJ
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Rodger, A., Marrington, R., Roper, D., Windsor, S. (2005). Circular Dichroism Spectroscopy for the Study of Protein-Ligand Interactions. In: Ulrich Nienhaus, G. (eds) Protein-Ligand Interactions. Methods in Molecular Biology, vol 305. Humana, Totowa, NJ. https://doi.org/10.1385/1-59259-912-5:343
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DOI: https://doi.org/10.1385/1-59259-912-5:343
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