Thermal dielectroscopy was used to study the spectrin-based sub-membrane skeleton (MS) and its attachment to the lipid bilayer of human erythrocyte membrane. At the spectrin denaturation temperature, T-A, the complex impedance, Z*, and capacitance, C*, sustain sigmoid, frequency-dependent changes, Delta Z* = Delta Z' + j Delta Z '' and Delta C* = Delta C' - j Delta C '', respectively. The impedance change plot, -Delta Z '' vs. Delta Z', depicts two semicircles revealing two dielectric relaxations on MS, here designated as beta and gamma relaxations on MS [Ivanov and Paarvanova, 2016]. We report here that ortho-vanadate (5 mM), alkaline cytosole (pH 9.2), N-ethylmaleimide (5 mM), diamide (1 mM) and cell shrinkage, all known to disrupt the band 3 tetramer-spectrin attachment site, inhibited gamma relaxation, which represents the direct interaction of high frequency field with spectrin. By contrast DNAase I, diphosphoglycerate (10 mM) and urea (1-2 M), all known to disrupt the actin-spectrin junction, inhibited beta relaxation, which represents the piezo effect on spectrin, powered by the low frequency electrostriction of lipid bilayer. The frequency curves of Delta C-d '' (dielectric component of Delta C '') and of Delta C' both determined the critical frequencies of beta and gamma relaxations as 1.4 and 6 MHz at 46 degrees C (1.1 and 4.5 MHz at 37 degrees C), respectively. Based on the presented results, the effect of MS attachment on the dynamics and elasticity of MS was discussed. (C) 2019 Elsevier Ltd. All rights reserved.