TY - JOUR

T1 - Cell-mediated reduction of protein and peptide hydroperoxides to reactive free radicals

AU - Headlam, Henrietta A

AU - Davies, Michael Jonathan

PY - 2003/1/1

Y1 - 2003/1/1

N2 - Radical attack on proteins in the presence of O(2) gives protein hydroperoxides in high yields. These peroxides are decomposed by transition metal ions, reducing agents, UV light and heat, with the formation of a range of reactive radicals that are capable of initiating further damage. Evidence has been presented for the formation of alcohols as stable products of peroxide decomposition, and these have been employed as markers of oxidative damage in vivo. The mechanism of formation of these alcohols is unclear, with both radical and nonradical pathways capable of generating these products. In this study we have investigated the reduction of peptide and protein hydroperoxides by THP-1 (human monocyte-like) cells and it is shown that this process is accompanied by radical formation as detected by EPR spin trapping. The radicals detected, which are similar to those detected from metal-ion catalyzed reduction, are generated externally to the cell. In the absence of cells, or with cell-conditioned media or cell lysates, lower concentrations of radicals were detected, indicating that intact cells are required for rapid hydroperoxide decomposition. The rate of radical generation was enhanced by preloading the cells with ascorbate, and this was accompanied by intracellular formation of the ascorbate radical. It is proposed that decomposition of some amino acid and peptide hydroperoxides occurs extracellularly via the involvement of a cell-surface reducing system, such as a trans-plasma membrane electron transport system (TPMET) either directly, or indirectly via redox cycling of trace transition metal ions.

AB - Radical attack on proteins in the presence of O(2) gives protein hydroperoxides in high yields. These peroxides are decomposed by transition metal ions, reducing agents, UV light and heat, with the formation of a range of reactive radicals that are capable of initiating further damage. Evidence has been presented for the formation of alcohols as stable products of peroxide decomposition, and these have been employed as markers of oxidative damage in vivo. The mechanism of formation of these alcohols is unclear, with both radical and nonradical pathways capable of generating these products. In this study we have investigated the reduction of peptide and protein hydroperoxides by THP-1 (human monocyte-like) cells and it is shown that this process is accompanied by radical formation as detected by EPR spin trapping. The radicals detected, which are similar to those detected from metal-ion catalyzed reduction, are generated externally to the cell. In the absence of cells, or with cell-conditioned media or cell lysates, lower concentrations of radicals were detected, indicating that intact cells are required for rapid hydroperoxide decomposition. The rate of radical generation was enhanced by preloading the cells with ascorbate, and this was accompanied by intracellular formation of the ascorbate radical. It is proposed that decomposition of some amino acid and peptide hydroperoxides occurs extracellularly via the involvement of a cell-surface reducing system, such as a trans-plasma membrane electron transport system (TPMET) either directly, or indirectly via redox cycling of trace transition metal ions.

KW - Cell Line

KW - Free Radicals

KW - Gamma Rays

KW - Humans

KW - Hydrogen Peroxide

KW - Oxidation-Reduction

KW - Peptides

KW - Proteins

M3 - Journal article

C2 - 12498978

VL - 34

SP - 44

EP - 55

JO - Free Radical Biology & Medicine

JF - Free Radical Biology & Medicine

SN - 0891-5849

IS - 1

ER -