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Updated: Oct 19, 2026

Fluorescent Leakage Assay to Investigate Membrane Destabilization by Cell-Penetrating Peptide
Published on: December 19, 2020
[Interaction of delta-sleep-inducing peptide with cell membranes in vitro]
G T Rikhireva1, I N Golubev, S A Kopylovskiĭ
1Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia.
Abstract:
The effect of delta-sleep-inducing peptide (DSIP) on erythrocytic membranes of human donor blood was studied by the spin label and spin probe methods. The spin-labeled derivative of DSIP containing the N-terminal residue of 1-oxyl-2,2,5,5-tetramethylpyrroline-3-carboxylic acid was synthesized. An analysis of the ESR spectra of the spin-labeled DSIP derivative recorded after its incubation with a human erythrocyte suspension at 37 degrees C revealed a decrease in the rotational correlation time (tau c) and molecular order parameter (S) in comparison with the control solutions of the peptide in phosphate buffer (pH 7.4). The application of paramagnetic probes, 5-, 12-, and 16-doxylstearic acids and 3-doxylandrostanol, demonstrated that the introduction of DSIP in an erythrocytic suspension significantly increased the mobility of the hydrophobic area of the membrane bilayer both at a depth of 20-22 A and in the subsurface area (4-6 A). The dependence of these effects on the DSIP concentration was shown to have the form of a curve with well-defined extremes. The maximal disordering of membrane lipids was observed at peptide concentrations of 10(-9) and 10(-6) M. These results suggested that DSIP significantly affected the structure of plasmatic membranes in vitro by changing the physical state of their lipid components.
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