Transient electric birefringence of human erythroid spectrin dimers and tetramers at ionic strengths of 4 mM and 53 mM

In conventional electrooptic studies the sample ionic strength must for technical reasons be kept below about 3 mM, which is only 2% of the ionic strength at physiological conditions. In particular for flexible polyelectrolytic macromolecules it can in general not be ruled out that both the conformational average and dynamics at ionic strength 3 mM and below may differ significantly from what it is at physiological conditions. Here we report on the first electrooptic study of human erythroid spectrin dimers and tetramers at ionic strengths higher than 3 mM. All measurements in this study were carried out at both ionic strength 4 mM (2.5 mM HEPES + 1 mM NaCl) and 53 mM (2.5 mM HEPES + 50 mM NaCl). Spectrin tetramers were studied only at 4 degrees C whereas the dimers were studied at both 4 degrees C and 37 degrees C. At 4 degrees C there is a striking quantitative similarity between the transient electric birefringence (TEB) of spectrin dimers and tetramers. Also, the TEB of spectrin dimers at 37 degrees C was very similar to the results at 4 degrees C. The contour length and the molecular weight of spectrin dimers and tetramers are known. The dominating TEB relaxation time is in all cases only a fraction of what is predicted theoretically if the spectrin dimers and tetramers are assumed to be stiff and extended molecules. In sum, the new TEB data constitute strong electrooptic evidence confirming that spectrin dimers and tetramers have a highly flexible structure, and demonstrate for the first time that a major part of the intrachain dynamics of the spectrin is quite insensitive to an increase of the ionic strength from 4 mM to 53 mM. Use of the reversing electric field pulse technique for all conditions studied yields TEB data suggesting that the orientation of both spectrin dimers and tetramers in an electric field is dominated by a permanent rather than an induced electric dipole moment.