Exp Mol Med.  1998 Dec;30(4):214-220.

Calyculin A modulates activation of the NADPH-oxidase in Me2SO-differentiated HL-60 cells

• Park JI
• Uhlinger D
• Chung BS
• Kim IH
• Kwak JY

Abstract

Human promyelocytic leukemia cells (HL-60) have been used as a model system in which to study the effects of protein phosphatase inhibitors on NADPH-oxidase activation. Since O2- is generated by NADPH-oxidase, we examined the effect of calyculin A pretreatment on oxidase activation in response to various agonists. When Me2SO-differentiated HL-60 cells were treated with calyculin A prior to the addition of phorbol 12-myristate 13-acetate (PMA), O2- production was inhibited; however, calyculin A enhanced O2- production by N-formyl-methionyl-leucyl-phenylalanine (FMLP). The decreased O2- production seen with calyculin A pretreatment followed by PMA may be due to diminished translocation of the p47-phox and p67-phox, cytosolic components of the oxidase, and inhibition of arachidonic acid release. Interestingly calyculin A pretreatment followed by either agonist significantly enhanced mitogen-activated-protein kinase (MAPK) activity. The differential effects of pretreatment with calyculin A on subsequent oxidase stimulation elicited by FMLP or PMA provide further evidence for substantial heterogeneity in the activation of the respiratory burst.

Keyword

NADPH-oxidase; calyculin A; p47-phox; p67-phox; arachidonic acid

MeSH Terms

Arachidonic Acid/metabolism
Ca(2+)-Calmodulin Dependent Protein Kinase/metabolism
Cell Differentiation
Dimethyl Sulfoxide/pharmacology*
Enzyme Inhibitors/pharmacology*
HL-60 Cells
Human
N-Formylmethionine Leucyl-Phenylalanine/pharmacology
NADPH Oxidase/metabolism*
Neutrophils/metabolism*
Neutrophils/drug effects
Oxazoles/pharmacology*
Oxygen/metabolism
Phosphoprotein Phosphatase/antagonists & inhibitors
Phosphoproteins/immunology
Signal Transduction
Tetradecanoylphorbol Acetate/pharmacology
Time Factors