The vitamin K-dependent carboxylation system in human osteosarcoma U2-OS cells. Antidotal effect of vitamin K1 and a novel mechanism for the action of warfarin.
An osteoblast-like human osteosarcoma cell line (U2-OS) has been shown to possess a vitamin K-dependent carboxylation system which is similar to the system in human HepG2 cells and in liver and lung from the rat. In an 'in vitro' system prepared from these cells, vitamin K1 was shown to overcome warfarin inhibition of gamma-carboxylation carried out by the vitamin K-dependent carboxylase. The data suggest that osteoblasts, the cells involved in synthesis of vitamin K-dependent proteins in bone, can use vitamin K1 as an antidote to warfarin poisoning if enough vitamin K1 can accumulate in the tissue. Five precursors of vitamin K-dependent proteins were identified in osteosarcoma and HepG2 cells respectively. In microsomes (microsomal fractions) from the osteosarcoma cells these precursors revealed apparent molecular masses of 85, 78, 56, 35 and 31 kDa. When osteosarcoma cells were cultured in the presence of warfarin, vitamin K-dependent 14C-labelling of the 78 kDa precursor was enhanced. Selective 14C-labelling of one precursor was also demonstrated in microsomes from HepG2 cells and from rat lung after warfarin treatment. In HepG2 cells this precursor was identified as the precursor of (clotting) Factor X. This unique 14C-labelling pattern of precursors of vitamin K-dependent proteins in microsomes from different cells and tissues reflects a new mechanism underlying the action of warfarin.
Wallin
R
R
Department of Medicine, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27103.
Rossi
F
F
Loeser
R
R
Key
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LL
D000931:Antidotes; D019735:Carbon-Carbon Ligases; D004591:Electrophoresis, Polyacrylamide Gel; D005170:Factor X / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008025:Ligases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008168:Lung / Q000378:metabolism / Q000648:ultrastructure; D008861:Microsomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D008970:Molecular Weight; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D012516:Osteosarcoma / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D011516:Prothrombin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014812:Vitamin K / Q000494:pharmacology*; D014859:Warfarin / Q000494:pharmacology* / Q000506:poisoning
D000931:Antidotes; D011498:Protein Precursors; D014812:Vitamin K; D014859:Warfarin; D011516:Prothrombin; D005170:Factor X; D008025:Ligases; D019735:Carbon-Carbon Ligases; C020660:glutamyl carboxylase