Benzimidazole Derivative NS1619 Inhibits Functioning of Mitochondria Isolated from Mouse Skeletal Muscle
M. V. Dubinin, A. D. Igoshkina, K. N. Belosludtsev
M. V. Dubinin, A. D. Igoshkina, K. N. Belosludtsev Biochemistry (Moscow), Supplement Series A: Membrane and Cell Biology. 2023;17(2):127-135
Pages: 127-135
Abstract
The surface potential of biological membranes, largely governed by the concentration of protons and other charged species at the lipid-water interface, is a critical determinant of membrane protein function and cellular signaling. This study investigates the utility of two widely-used electrochromic styryl dyes, di-4-ANEPPS and RH-421, not merely as voltage-sensitive probes, but specifically as sensors for protons on the surface of model lipid membranes. The research objective was to characterize the pH-dependent changes in the optical properties of these dyes when adsorbed onto bilayer lipid membranes (BLMs) and liposomes. Using fluorescence spectroscopy and measurements of the boundary potential ($\Delta\varphi_b$), we demonstrate that the fluorescence intensity and spectral shift of both di-4-ANEPPS and RH-421 are highly sensitive to the local proton concentration. This sensitivity is attributed to the protonation/deprotonation of the dye molecules or nearby lipid headgroups, which alters the dipole potential at the membrane interface. The findings confirm that these styryl dyes can serve as effective, real-time optical reporters for proton binding and surface charge dynamics, providing a valuable tool for studying proton-coupled processes and local pH microenvironments in cell biology and biophysics.