Back to Search Start Over

Spontaneous intermembrane transfer of various cholesterol-derived hydroperoxide species : kinetic studies with model membranes and cells

Authors :
Albert W. Girotti
Witold Korytowski
Andrew Vila
Publication Year :
2001

Abstract

Whereas spontaneous and protein-mediated transfer/exchange of cholesterol (Ch) between membranes has been widely studied, relatively little is known about the translocation of Ch oxidation products, particularly hydroperoxide species (ChOOHs), which can act as cytotoxic prooxidants. A major aim of the present study was to examine and compare the intermembrane transfer characteristics of several biologically relevant ChOOH isomers, including singlet oxygen-derived 5alpha-OOH, 6alpha-OOH, and 6beta-OOH and free radical-derived 7alpha-OOH and 7beta-OOH. These species were generated in [(14)C]Ch-labeled donor membranes [erythrocyte ghosts or unilamellar DMPC/Ch (1.0:0.8 mol/mol) liposomes] by means of dye-sensitized photoperoxidation. Spontaneous transfer to nonoxidized acceptor membranes (DMPC liposomes or ghosts, respectively) at 37 degrees C was monitored by thin-layer chromatography with phosphorimaging radiodetection (HPTLC-PI) or liquid chromatography with mercury cathode electrochemical detection [HPLC-EC(Hg)]. The former allowed measurement of total (unresolved) ChOOH along with parent Ch, whereas the latter allowed measurement of individual ChOOHs. Ghost membranes in which approximately 4% of the Ch had been peroxidized, giving mainly 5alpha-OOH, transferred total ChOOH and Ch to liposomes in apparent first-order fashion, the rate constant for ChOOH being approximately 65 times greater. Like Ch desorption, ChOOH desorption from donor membranes was found to be rate limiting, and rate varied inversely with size when liposomal donors were used. For individual ChOOHs, rate constant magnitude (7alpha/7beta-OOH > 5alpha-OOH > 6alpha-OOH > 6beta-OOH) correlated inversely with reverse-phase HPLC retention time, suggesting that faster transfer reflects greater hydrophilicity. Liposome-borne ChOOHs exhibited the same order of toxicity toward COH-BR1 cells, which are deficient in ability to detoxify these peroxides. The prospect of disseminating oxidative cell injury via translocation of ChOOHs and other lipid hydroperoxides is readily apparent from these findings.

Details

Language :
English
Database :
OpenAIRE
Accession number :
edsair.doi.dedup.....74a09e8ead42f8d57b8c27774afc2e0b