1. Summary
The NADPH oxidase producing-superoxide is the major mechanism by which phagocytes kill invading pathogens. The human myeloid cell line PLB-985 was transfected to express p85 cytosolic phospholipase A2 (cPLA2) antisense mRNA and stable clones were selected which lack detectable cPLA2. cPLA2-deficient PLB-985 cells differentiate similarly to control PLB-985 cells in response to retinoic acid, DMSO or 1,25 dihydroxyvitamin D3 indicating that cPLA2 is not involved in the differentiation process. Despite the normal synthesis of NADPH oxidase subunits during differentiation of cPLA2-deficient PLB-985 cells, these cells fail to activate NADPH oxidase in response to a variety of soluble and particulate stimuli, but addition of exogenous arachidonic acid (AA) fully restores oxidase activity. This establishes an essential requirement of cPLA2 generated AA for activation of phagocyte NADPH oxidase. In order to elucidate the mechanism by which cPLA2 regulates the oxidase, the role of cPLA2 in NADPH oxidase associated H+ channel was studied. Activation of differentiated PLB cells resulted in a Zn+2 sensitive alkalization, indicating H+ channel activity. In contrast, differentiated PLB-D cells failed to activate the H+ channel, but addition of exogenous AA fully restored this activity, indicating an essential and specific physiological requirement of cPLA2-generated AA for activation of the H+ channel. The presence of the H+ channel inhibitor, Zn+2, caused significant inhibition of NADPH oxidase activity, suggesting a role of the NADPH oxidase associated H+ channel in regulating oxidase activity.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Babior, B. M. (1999). NADPH oxidase. an update. Blood 93:1464–76.
Bolognese, B., McCord, M., and Marshall, L. A. (1995). Differential regulation of elicited-peritoneal macrophage 14 kDa and 85 kDa phospholipase A2(s) by transforming growth factor-beta. Biochim Biophys Acta 1256:201–9.
Boron, W. F. (1983). Transport of H+ and of ionic weak acids and bases. JMembr Biol 72: 1–16.
Clark, J. D., Milona, N., and Knopf, J. L. (1990). Purification of a 110-kilodalton cytosolic phospholipase A2 from the human monocytic cell line U937. Proc Nutl Acad Sci USA 87:7708–12.
Cockcroft, S. (1991). Relationship between arachidonate release and exocytosis in permeabilized human neutrophils stimulated with formylmethionyl-leucyl-phenylalanine (fMetLeuPhe), guanosine 5′-[gamma-thio]triphosphate (GTP[S]) and Ca2+. Biochem J. 275: 127–31.
Hazan, I., Dana, R., Granot, Y., and Levy, R. (1997). Cytosolic phospholipase A, and its mode of activation in human neutrophils by opsonized zymosan. Correlation between 42/44 kDa mitogen-activated protein kinase, cytosolic phospholipase A2 and NADPH oxidase. Biochem J. 326: 867–76.
Hazen, S. L., Hall, C. R., Ford, D. A., and Gross, R. W. (1993). Isolation of a human myocardial cytosolic phospholipase A2 isoform. Fast atom bombardment mass spectroscopic and reverse-phase high pressure liquid chromatography identification of choline and ethanolamine glycerophospholipid substrates. J Clin Invest 91.
Henderson, L. M. (1998). Role of histidines identified by mutagenesis in the NADPH oxidase-associated H+ channel. J Biol Chem. 273:33216–23.
Henderson, L. M., Banting, G., and Chappell, J. B. (1995). The arachidonate-activable, NADPH oxidase-associated H+ channel. Evidence that gp91-phox functions as an essential part of the channel. J Biol Chem. 270:5909–16.
Henderson, L. M., and Chappell, J. B. (1992). The NADPH-oxidase-associated H+ channel is opened by arachidonate. Biochem 5.283: 171–5.
Henderson, L. M., Chappell, J. B., and Jones, 0. T. (1987). The superoxide-generating NADPH oxidase of human neutrophils is electrogenic and associated with an H+ channel. Biochem J 246, 325–9.
Henderson, L. M., Thomas, S., Banting, G., and Chappell, J. B. (1997). The arachidonate-activatable, NADPH oxidase-associated H+ channel is contained within the multi-membrane-spanning N-terminal region of gp91-phox. Biochem J 325, 701–5.
Kapus, A., Romanek, R., and Grinstein, S. (1994). Arachidonic acid stimulates the plasma membrane H+ conductance of macrophages. J Biol Chem 269, 4736–45.
Kapus, A., Susztak, K., and Ligeti, E. (1993). Regulation of the electrogenic H+ channel in the plasma membrane of neutrophils: possible role of phospholipase A2, internal and external protons. Biochem J 292, 445–50.
Kramer, R. M., Hession, C., Johansen, B., Hayes, G., McGray, P., Chow, E. P., Tizard, R., and Pepinsky, R. B. (1989). Structure and properties of a human non-pancreatic phospholipase A2. J Bioi Chem 264, 5768–75.
Lefkowith, J. B., Lennartz, M. R., Rogers, M., Morrison, A. R., and Brown, E. J. (1992). Phospholipase activation during monocyte adherence and spreading. J lmmunol 149, 1729–35.
Lennartz, M. R., and Brown, E. J. (1991). Arachidonic acid is essential for IgG Fc receptor-mediated phagocytosis by human monocytes. J Immunol 147, 621–6.
Nanda, A., and Grinstein, S. (1991). Protein kinase C activates an H+ (equivalent) conductance in the plasma membrane of human neutrophils. Proc Nutl Acad Sci USA 88, 10816–20.
Rosenthal, M. D., Gordon, M. N., Buescher, E. S., Slusser, J. H., Harris, L. K., and Franson, R. C. (1995). Human neutrophils store type II 14-kDa phospholipase A, in granules and secrete active enzyme in response to soluble stimuli. Biochem Biophys Res Commun 208, 650–6.
Rotrosen, D., Yeung, C. L., Leto, T. L., Malech, H. L., and Kwong, C. H. (1992). Cytochrome b558: the flavin-binding component of the phagocyte NADPH oxidase. Science 256, 1459–62.
Seilhamer, J. J., Pruzanski, W., Vadas, P., Plant, S., Miller, J. A., Kloss, J., and Johnson, L. K. (1989). Cloning and recombinant expression of phospholipase A, present in rheumatoid arthritic synovial fluid. J Bioi Chem 264, 5335–8.
Susztak, K., Mocsai, A., Ligeti, E., and Kapus, A. (1997). Electrogenic H+ pathway contributes to stimulus-induced changes of internal pH and membrane potential in intact neutrophils: role of cytoplasmic phospholipase A2. Biochem J 325, 501–10.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Kluwer Academic Publishers
About this chapter
Cite this chapter
Levy, R., Lowenthal, A., Dana, R. (2002). Cytosolic Phospholipase A2 is Required for the Activation of the Nadph Oxidase Associated H+ Channel in Phagocyte-Like Cells. In: Keisari, Y., Ofek, I. (eds) The Biology and Pathology of Innate Immunity Mechanisms. Advances in Experimental Medicine and Biology, vol 479. Springer, Boston, MA. https://doi.org/10.1007/0-306-46831-X_11
Download citation
DOI: https://doi.org/10.1007/0-306-46831-X_11
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-306-46409-6
Online ISBN: 978-0-306-46831-5
eBook Packages: Springer Book Archive