Electrooptic measurement procedure

A sensitive approach for the analysis of molecular structures is the measurement of rotation time constants and optical anisotropy coefficients by relaxation electrooptical methods.The experimental procedure used for electrooptical investigations is relatively simple samples are subjected to electric field pulses, and the response due to field-induced alignment or field-induced reactions is recorded by spectrophotometric techniques. In the dichroism experiments, the absorbance of polarized light is measured under electric field pulses. The measured quantity, linear dichroism (LD), means that anisotropic absorption of plane or linearly polarized light has taken place. 

Experimental dipole moments may be used to check the validity of electrostatic calculations. In the past, dipole moments of proteins were often characterized by measurements of dielectric relaxation. More information may be obtained by measurements of the electric dichroism because these measurements provide not only the magnitude of the dipole moment but also the optical anisotropy with respect to the dipole vector. Thus, measurements of the electric dichroism provide a more rigorous test for calculations of electrostatic parameters of proteins. Using the calculations described earlier for pK s of titratable groups, one can predict dipole moments of proteins and their axes given by the principal axes of the rotational diffusion tensor and compare them with electrooptical data. ° One important aspect of comparison of computed and experimental dipole moment is that computations of dipole moments, optical anisotropy, and rotational diffusion coefficients can be used in combination with experimental electrooptical procedures to determine the long-range structure of biomacromolecular assemblies, such as the complexes of DNA and proteins described by Pbrschke et al. so... 

---