Reanalysis of the testosterone displacement from its HSA binding site by DHEA using competitive Langmuir isotherm

Abstract

In a previous manuscript [C. Andre, A. Berthelot, J.F. Robert, M. Thomassin, Y.C. Guillaume, J. Pharm. Bio. Med., in press], the Mg 2+ effect on the testosterone displacement equilibrium from its human serum albumin (HSA) binding site by DHEA was investigated using a thermodynamic approach. In this paper, a novel concept based on the competitive Langmuir distribution isotherms was proposed to calculate the association constant of the HSA–hormone binding and to confirm the Mg 2+ role on the testosterone displacement equilibrium from its HSA binding site by DHEA. Thus, both the HSA–hormone binding and the displacement equilibrium processes were reanalysed. The results obtained confirmed that: – DHEA and testosterone bound on the same HSA site; – DHEA displaced well testosterone to its HSA binding site; – Mg 2+ cation led a change on both the HSA–hormone binding and the displacement equilibrium at a critical Mg 2+ concentration x c around 1 mM.