The Effect of Highly Hydroxylated Fullerenol C₆₀(OH)₃₆on Human Erythrocyte Membrane Organization

Abstract

The mechanism of the interaction of highly hydroxylated fullerenol C60(OH)36with erythrocyte membranes was studied by electron spin resonance spectroscopy (ESR) of stearic acid derivatives labeled with a nitroxyl radical at C-12 or C-16 and with a nitroxyl derivative of maleimide covalently attached to sulfhydryl groups of membrane proteins. A significant increase in membrane fluidity in the hydrophobic region of the lipid bilayer was observed for 12-doxylstearic acid at fullerenol concentrations of 100 mg/L or 150 mg/L, while for 16-doxylstearic acid significant increase in fluidity was only observed at 150 mg/L. Fullerenol at 100 mg/L or 150 mg/L caused conformational changes in membrane proteins, expressed as an increase in thehw/hsparameter, when fullerenol was added before the maleimide spin label (MSL) to the membrane suspension. The increase of thehw/hsparameter may be caused by changes in lipid-protein or protein-protein interactions which increase the mobility of the MSL label and as a result increase the membrane fluidity. Incubation of the membranes with the MSL before the addition of fullerenol blocked the available membrane protein –SH groups and minimized the interaction of fullerenol with them. This confirms that fullerenol interacts with erythrocyte membrane proteins via available protein –SH groups.

Current advances in nanomaterials affecting morphology, structure, and function of erythrocytes

“…In addition to their intrinsic properties, NPs are also inuenced by a variety of external factors, including the type of culture medium, erythrocyte species, concentration, storage duration, animal health status, centrifugation rate, exposure time, and incubation temperature. 8 These studies have offered evidence that in the case of GOrelated NPs, the high extent of exfoliation of the material, the 11,25,34,38,39,45…”

“…Effects of various nanoparticles on oxygen-carrying functions, longevity and membrane oxidative stress of erythrocytes NPs 11,25,34,38,39,45 Prevent its degradation…”

“…38 They also found that fullerenol C 60 (OH) 36 could interact with functional sites on erythrocyte membranes with available -SH groups to retain membrane -SH concentration, increase h W /h S parameters, and stabilize membrane protein conformation to maintain membrane protein structure and function. 11,39 Oxidative stress can induce band 3 protein rearrangement on the erythrocyte membrane, and under repeated oxidative perturbations, band 3 clusters progressively grow and shi to an irreversible state. 40 Studies have shown that phagocytic recognition of senescent and oxidatively stressed erythrocytes also appears to be initiated by band 3 clustering, accompanied by bivalent binding of anti-band 3 autoantibodies and oligomerized band 3.…”

Carbon Nanomaterials Alter Global Gene Expression Profiles

“…However, a recent study has definitively confirmed that highly hydroxylated fullerenol, C 60 (OH) 36 , are able to penetrate the hydrophobic core of the lipid bilayer of red blood cells, in vitro. 57 In addition, the authors observed that C 60 (OH) 36 interacted with available SH groups of erythrocyte membrane proteins, accompanied by conformational membrane changes and a decrease in membrane fluidity. 57 We reasonably expect similar interactions of C 60 (OH) 24 with the plasma membrane of yeast cells, though experimental confirmation of this relationship in live systems should be followed.…”

“…57 In addition, the authors observed that C 60 (OH) 36 interacted with available SH groups of erythrocyte membrane proteins, accompanied by conformational membrane changes and a decrease in membrane fluidity. 57 We reasonably expect similar interactions of C 60 (OH) 24 with the plasma membrane of yeast cells, though experimental confirmation of this relationship in live systems should be followed. This interaction at the membrane level also makes it possible for single or clusters of fullerenols to enter the cytosol, possibly through endocytosis.…”