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• W2273763634 abstract "Extracellular and intracellular reactive oxygen species attack different targets and may, therefore, result in different forms of oxidative stress. To specifically study an oxidative stress induced by a regulated intracellular flux of a defined reactive oxygen species in endothelium, we used immunotargeting of the H 2 O 2 -generating enzyme glucose oxidase (GOX) conjugated with an antibody to platelet-endothelial cell adhesion molecule (PECAM)-1, an endothelial surface antigen. Anti-PECAM- 125 I-GOX conjugates specifically bind to both endothelial and PECAM-transfected cells. Approximately 70% of cell-bound anti-PECAM- 125 I-GOX was internalized. The cell-bound conjugate was enzymatically active and generated H 2 O 2 from glucose. Use of the fluorescent dye dihydrorhodamine 123 revealed that 70% of H 2 O 2 was generated intracellularly, whereas 30% of H 2 O 2 was detected in the cell medium. Catalase added to the cells eliminated H 2 O 2 in the medium but had little effect on the intracellular generation of H 2 O 2 by anti-PECAM-GOX. Both H 2 O 2 added exogenously to the cell medium (extracellular H 2 O 2 ) and that generated by anti-PECAM-GOX caused oxidative stress manifested by time- and dose-dependent irreversible plasma membrane damage. Inactivation of cellular catalase by aminotriazole treatment augmented damage caused by either extracellular H 2 O 2 or anti-PECAM-GOX. Catalase added to the medium protected either normal or aminotriazole-treated cells against extracellular H 2 O 2 , yet failed to protect cells against injury induced by anti-PECAM-GOX. Therefore, treatment of PECAM-positive cells with anti-PECAM-GOX leads to conjugate internalization, predominantly intracellular H 2 O 2 generation and intracellular oxidative stress. These results indicate that anti-PECAM-GOX 1) provides cell-specific intracellular delivery of an active enzyme and 2) causes intracellular oxidative stress in PECAM-positive cells." @default.
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• W2273763634 date "1999-08-01" @default.
• W2273763634 modified "2023-10-14" @default.
• W2273763634 title "Immunotargeting of glucose oxidase: intracellular production of H₂O₂and endothelial oxidative stress" @default.
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• W2273763634 doi "https://doi.org/10.1152/ajplung.1999.277.2.l271" @default.
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