Transplasma membrane redox system of HL-60 cells is controlled by cAMP.
Transplasma membrane redox activity of HL-60 cells was determined by measuring the prevention of ascorbate chemical oxidation. The ascorbate free radical produced as the first step of ascorbate oxidation was reduced back by the transplasma membrane electron transport system, causing then the regeneration of extracellular ascorbate. Agents that increase intracellular cAMP, such as forskolin and dibutyryl cAMP (db-cAMP), increased the rate of ascorbate regeneration by HL-60 cells. Also, the phosphodiesterase-resistant cAMP analogue Sp-cAMP-S (agonist of the protein kinase A) increased the electron flow to the ascorbate free radical at the plasma membrane. Rp-cAMP-S, antagonist of the protein kinase A, partially inhibited the redox activity of cells and abolished the effect of Sp-cAMP-S. Inhibition obtained after preincubation of cells in Rp-cAMP-S was reversed by Sp-cAMP-S. Tunicamycin, a compound that inhibited the electron flow to the ascorbate free radical at the plasma membrane, also reduced the response of transplasma membrane redox system to Sp-cAMP-S. Lactate slightly affected the ascorbate regeneration in nonstimulated cells, but showed a significant effect on Sp-cAMP-S-stimulated plasma membrane electron flow. We show here a role for cAMP in the short-term modulation of transplasma membrane redox system measured as the regeneration of ascorbate at the cell surface of HL-60 cells, probably mediated by cAMP-dependent protein kinases.
Rodríguez-Aguilera
J C
JC
Departamento de Biología Celular, Facultad de Ciencias, Universidad de Córdoba, Spain.
Nakayama
K
K
Arroyo
A
A
Villalba
J M
JM
Navas
P
P
D001205:Ascorbic Acid / Q000378:metabolism; D003994:Bucladesine / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology*; D004579:Electron Transport; D007773:Lactates / Q000494:pharmacology; D019344:Lactic Acid; D010084:Oxidation-Reduction; D014415:Tunicamycin / Q000494:pharmacology
D007773:Lactates; D014415:Tunicamycin; D005576:Colforsin; D019344:Lactic Acid; D003994:Bucladesine; D000242:Cyclic AMP; D001205:Ascorbic Acid