Regulation of Cu,Zn superoxide dismutase with copper. Caeruloplasmin maintains levels of functional enzyme activity during differentiation of K562 cells.
K562 cells, a human erythroleukaemic cell line blocked for differentiation, commit towards erythrocytes when exposed to haemin (20 microM). The cells synthesize fetal haemoglobins and show site-specific binding of caeruloplasmin, a plasma copper protein. These events are set into motion by haemin. On the assumption that the binding of caeruloplasmin could reflect a greater need for copper, we sought to determine whether the transfer of 67Cu from caeruloplasmin was accelerated in haemin-induced compared with non-induced K562 cells. Cu,Zn superoxide dismutase (CuZnSOD) was the recipient. Haemin induction caused the K562 cells to lose CuZnSOD activity. By 96 h, the level of SOD activity was less than 60% of that of non-induced cells. The loss was confined entirely to the CuZn form, MnSOD activity staying essentially unchanged. Although CuZnSOD activity declined with the haemin induction, the incorporation of [4,5-3H]lysine into immunoprecipitable CuZnSOD protein was unaffected. There was also no change in CuZnSOD mRNA concentration in haemin-induced cells. Thus a loss of enzyme did not correlate with a decline in the synthesis de novo of CuZnSOD protein. When 48 h-induced cells were transferred to a medium supplemented with 0.2 microM-caeruloplasmin, CuZnSOD activity was restored to control levels in 24 h. Caeruloplasmin also stimulated the incorporation of [3H]lysine into immunoprecipitable CuZnSOD protein. Caeruloplasmin addition may have affected a post-translational regulatory site for CuZnSOD biosynthesis, possibly by providing copper for the newly synthesized enzyme.
Percival
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Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843.
Harris
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D001692:Biological Transport; D002454:Cell Differentiation; D002460:Cell Line; D002570:Ceruloplasmin / Q000494:pharmacology*; D003300:Copper / Q000378:metabolism / Q000494:pharmacology*; D004261:DNA Replication / Q000187:drug effects; D006418:Heme / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute; D008239:Lysine / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000378:metabolism; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*
D009363:Neoplasm Proteins; D006418:Heme; D003300:Copper; D013482:Superoxide Dismutase; D002570:Ceruloplasmin; D008239:Lysine