HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Published online before print December 9, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: jlb.0504284v1 77/3/414    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zhao, X. Articles by Cathcart, M. K. Search for Related Content PubMed PubMed Citation Articles by Zhao, X. Articles by Cathcart, M. K.

(Journal of Leukocyte Biology. 2005;77:414-420.)
© 2005 by Society for Leukocyte Biology

Protein kinase C regulates p67phox phosphorylation in human monocytes

Xiaoxian Zhao^*, Bo Xu^*, Ashish Bhattacharjee^*, Claudine M. Oldfield^*, Frans B. Wientjes, Gerald M. Feldman and Martha K. Cathcart^*,1

^* Department of Cell Biology, Lerner Research Institute, Cleveland Clinic Foundation, Ohio;
Department of Medicine, University College London, United Kingdom; and
Division of Monoclonal Antibodies, Office of Therapeutics, Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland

¹ Correspondence: Department of Cell Biology, Lerner Research Institute, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195. E-mail: cathcam{at}ccf.org

Phosphorylation of the reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase components p67phox and p47phox accompanies the assembly and activation of this enzyme complex. We have previously reported that activation of human monocytes with opsonized zymosan (ZOP), a potent stimulator of NADPH oxidase activity, results in the phosphorylation of p67phox and p47phox. In this study, we investigated the regulation of p67phox phosphorylation. Although protein kinase C (PKC) has previously been shown to regulate NADPH oxidase activity, we found that inhibition of PKC had no effect on p67phox phosphorylation. Our studies demonstrate that pretreatment of monocytes with antisense oligodeoxyribonucleotides specific for PKC or rottlerin, a selective inhibitor for PKC, inhibited the phosphorylation of p67phox in monocytes, and Go6976, a specific inhibitor for conventional PKCs, PKC and PKCß, had no such inhibitory effect. Additional studies indicate that ZOP stimulation of monocytes induces PKC and p67phox to form a complex. We also demonstrate that lysates from activated monocytes as well as PKC immunoprecipitates from activated monocytes can phosphorylate p67phox in vitro and that pretreatment of monocytes with rottlerin blocked the phosphorylation in each case. We further show that recombinant PKC can phosphorylate p67phox in vitro. Finally, we show that PKC-deficient monocytes produce significantly less superoxide anion in response to ZOP stimulation, thus emphasizing the functional significance of the PKC regulation of p67phox phosphorylation. Taken together, this is the first report to describe the requirement of PKC in regulating the phosphorylation of p67phox and the related NADPH oxidase activity in primary human monocytes.

Key Words: PKC • NADPH oxidase • superoxide anion • inflammation

This article has been cited by other articles:

				N. J. D. McLaughlin, A. Banerjee, S. Y. Khan, J. L. Lieber, M. R. Kelher, F. Gamboni-Robertson, F. R. Sheppard, E. E. Moore, G. W. Mierau, D. J. Elzi, et al. Platelet-Activating Factor-Mediated Endosome Formation Causes Membrane Translocation of p67phox and p40phox That Requires Recruitment and Activation of p38 MAPK, Rab5a, and Phosphatidylinositol 3-Kinase in Human Neutrophils J. Immunol., June 15, 2008; 180(12): 8192 - 8203. [Abstract] [Full Text] [PDF]

				A. Schwegmann, R. Guler, A. J. Cutler, B. Arendse, W. G. C. Horsnell, A. Flemming, A. H. Kottmann, G. Ryan, W. Hide, M. Leitges, et al. Protein kinase C {delta} is essential for optimal macrophage-mediated phagosomal containment of Listeria monocytogenes PNAS, October 9, 2007; 104(41): 16251 - 16256. [Abstract] [Full Text] [PDF]

				L. T. Huang, C. J. Paredes, E. T. Papoutsakis, and W. M. Miller Gene expression analysis illuminates the transcriptional programs underlying the functional activity of ex vivo-expanded granulocytes Physiol Genomics, September 11, 2007; 31(1): 114 - 125. [Abstract] [Full Text] [PDF]

				K. Bedard and K.-H. Krause The NOX Family of ROS-Generating NADPH Oxidases: Physiology and Pathophysiology Physiol Rev, January 1, 2007; 87(1): 245 - 313. [Abstract] [Full Text] [PDF]

				I. Kuwahara, E. P. Lillehoj, W. Lu, I. S. Singh, Y. Isohama, T. Miyata, and K. C. Kim Neutrophil elastase induces IL-8 gene transcription and protein release through p38/NF-{kappa}B activation via EGFR transactivation in a lung epithelial cell line Am J Physiol Lung Cell Mol Physiol, September 1, 2006; 291(3): L407 - L416. [Abstract] [Full Text] [PDF]

				V. Thakur, M. T. Pritchard, M. R. McMullen, Q. Wang, and L. E. Nagy Chronic ethanol feeding increases activation of NADPH oxidase by lipopolysaccharide in rat Kupffer cells: role of increased reactive oxygen in LPS-stimulated ERK1/2 activation and TNF-{alpha} production J. Leukoc. Biol., June 1, 2006; 79(6): 1348 - 1356. [Abstract] [Full Text] [PDF]

				M. Oda, S. Ikari, T. Matsuno, Y. Morimune, M. Nagahama, and J. Sakurai Signal Transduction Mechanism Involved in Clostridium perfringens Alpha-Toxin-Induced Superoxide Anion Generation in Rabbit Neutrophils. Infect. Immun., May 1, 2006; 74(5): 2876 - 2886. [Abstract] [Full Text] [PDF]