Increase of Cu,Zn-superoxide dismutase activity during differentiation of human K562 cells involves activation by copper of a constantly expressed copper-deficient protein.
Cu,Zn-superoxide dismutase activity, expressed on the basis of cell number, increased by 50% during sodium butyrate-induced differentiation of human K562 erythroleukemia cells. The increased enzyme activity was found to be concomitant with constant Cu,Zn-superoxide dismutase mRNA and immunoreactive protein levels and was accompanied by a rise in intracellular copper and glutathione. Incubation of K562 cell homogenates with copper caused an increase of Cu,Zn-superoxide dismutase activity which reached the levels observed after differentiation in the presence of sodium butyrate. The same treatment led to no significant activity increase in homogenates derived from differentiated cells. Externally added ceruloplasmin increased both intracellular copper levels and Cu,Zn-superoxide dismutase activity in undifferentiated cells to a level comparable with that observed after induction of differentiation. Both increments were abolished by depletion of cell glutathione. Cu,Zn-superoxide dismutase purified from control cells had both a lower kcat and a lower copper content than the enzyme purified from differentiated cells. From these data we conclude that: 1) Cu,Zn-superoxide dismutase is present in K562 cells also under the form of a less active copper-deficient enzyme, 2) the extent of enzyme activation is regulated post-translationally by differential delivery of copper as a function of differentiation stage, and 3) glutathione is likely to play a role in delivering copper to the copper-deficient protein in intact K562 cells.
Steinkühler
C
C
Department of Biology, University of Rome, Tor Vergata, Italy.
Sapora
O
O
Carrì
M T
MT
Nagel
W
W
Marcocci
L
L
Ciriolo
M R
MR
Weser
U
U
Rotilio
G
G
D002374:Catalase / Q000378:metabolism; D002454:Cell Differentiation; D002570:Ceruloplasmin / Q000378:metabolism; D002850:Chromatography, Gel; D003300:Copper / Q000172:deficiency / Q000378:metabolism*; D004789:Enzyme Activation; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute; D008668:Metallothionein / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D003300:Copper; D008668:Metallothionein; D002374:Catalase; D013482:Superoxide Dismutase; D002570:Ceruloplasmin; D005978:Glutathione