Redox modulation of protein kinase/phosphatase balance in melanoma cells: the role of endogenous and gamma-glutamyltransferase-dependent H2O2 production.
Alterations of protein kinase and protein phosphatase activities have been described in a number of tumors. Redox changes, such as in conditions of oxidant stress, have been reported to affect the cellular protein kinase/phosphatase balance. A basal production of reactive oxygen species (ROS), such as hydrogen peroxide (H(2)O(2)), exists in tumor cells, and the membrane-bound ecto-enzyme gamma-glutamyltransferase (GGT)-overexpressed in a variety of malignant tumors-is one of the mechanisms capable of promoting such a production. The present study was aimed to verify the interactions of GGT activity with protein phosphatase and kinase activities in Me665/2/60 melanoma cells, expressing high levels of this enzyme and exhibiting both basal and GGT-dependent production of hydrogen peroxide. An increase of total phosphatase as well as tyrosine phosphatase activities was observed after treatment of cells with both micromolar H(2)O(2) and GGT stimulation. Accordingly, stimulation of GGT resulted in decreased levels of phosphotyrosine. On the other hand, when serine/threonine phosphatase activities were selectively analyzed, both H(2)O(2) treatment and GGT stimulation caused their down-regulation.The data reported suggest that basal conditions of oxidant stress in melanoma may represent a factor contributing to the redox regulation of protein phosphorylation, and that GGT-mediated prooxidant reactions may participate in the process. As basal oxidant stress and expression of GGT activity are present in a variety of malignant tumors besides melanoma, these phenomena likely represent general mechanisms participating in the alteration of intracellular transduction during carcinogenesis.
Pieri
Lisa
L
Dip. di Patologia Sperimentale and B.M.I.E., Sez Pathologia Generale and Clinica, Scuola Medica, Università di Pisa, Via Roma 55, 56126, Pisa, Italy.
Dominici
Silvia
S
Del Bello
Barbara
B
Maellaro
Emilia
E
Comporti
Mario
M
Paolicchi
Aldo
A
Pompella
Alfonso
A
D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism*; D008545:Melanoma / Q000201:enzymology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010749:Phosphoprotein Phosphatases / Q000032:analysis / Q000378:metabolism*; D019000:Phosphotyrosine / Q000032:analysis; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000032:analysis; D017027:Protein Tyrosine Phosphatases / Q000032:analysis; D011505:Protein-Tyrosine Kinases / Q000032:analysis; D014407:Tumor Cells, Cultured; D005723:gamma-Glutamyltransferase / Q000096:biosynthesis / Q000378:metabolism*
D019000:Phosphotyrosine; D006861:Hydrogen Peroxide; D005723:gamma-Glutamyltransferase; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D010749:Phosphoprotein Phosphatases; D017027:Protein Tyrosine Phosphatases