Effects of deferoximine on chondrocyte alkaline phosphatase activity: proxidant role of deferoximine in thalassemia.
The homozygous form of beta-thalassemia, the most common single gene disorder, is treated by red cell transfusion therapy. Following transfusion, the chelator, deferoximine, is administered to patients to remove excess iron. However, when this drug is given to young children, metaphyseal dysplasia and abnormalities of linear growth are frequently observed. To explore the notion that deferoximine interferes with endochondral growth by chelating zinc, we examined the effect of the drug on chondrocytes maintained in long-term culture. We found that deferoximine caused a dose-dependent inhibition of a wide range of functions including cell proliferation, protein synthesis (and possibly under-hydroxylation of type X collagen), and mineral deposition. Directly relevant to the mineralization process was the observation that the drug dramatically lowered the activity of alkaline phosphatase, a zinc-requiring enzyme. To test the hypothesis that enzyme inhibition was due to chelation of zinc by deferoximine, the cell culture medium was supplemented with excess zinc. However, this treatment did not overcome the deferoximine-dependent change in enzyme activity. We next examined the possibility that deferoximine, in the presence of ascorbate, could form a free radical system that would serve to inactivate the enzyme. Using alkaline phosphatase extracted from chick cartilage, we noted that the activity of the phosphatase was markedly reduced in the presence of deferoximine and ascorbate. These effects were consistant with the notion that deferoximine and ascorbate can act as a prooxidant couple. This conclusion was confirmed when we measured the oxidative activities of the system using nitrobule tetrazolium and cytochrome c. Indeed, we noted that deferoximine markedly activates the autocatalytic oxidation of ascorbate.(ABSTRACT TRUNCATED AT 250 WORDS)
Hatori
M
M
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104-6003, USA.
Sparkman
J
J
Teixeira
C C
CC
Grynpas
M
M
Nervina
J
J
Olivieri
N
N
Shapiro
I M
IM
D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D002356:Cartilage / Q000187:drug effects* / Q000201:enzymology; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology*; D002642:Chick Embryo; D003676:Deferoxamine / Q000494:pharmacology*; D015394:Molecular Structure; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D015032:Zinc / Q000378:metabolism*; D017086:beta-Thalassemia / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology
D002614:Chelating Agents; D011500:Protein Synthesis Inhibitors; D000469:Alkaline Phosphatase; D003676:Deferoxamine; D015032:Zinc