Knockout of p47 phox Uncovers a Critical Role of p40 phox in Reactive Oxygen Species Production in Microvascular Endothelial Cells
Open accessAbstract
Objective— p40 phox is an important regulatory subunit of NADPH oxidase, but its role in endothelial reactive oxygen species (ROS) production remains unknown. Methods and Results— Using coronary microvascular endothelial cells isolated from wild-type and p47 phox knockout mice, we found that knockout of p47 phox increased the level of p40 phox expression, whereas depletion of p40 phox in wild-type cells increased p47 phox expression. In both cases, the basal ROS production (without agonist stimulation) was well preserved. Double knockout of p40 phox and p47 phox dramatically reduced (≈65%) ROS production and cells started to die. The transcriptional regulation of p40 phox and p47 phox expressions involves HBP1. p40 phox was prephosphorylated in resting cells. PMA stimulation induced p40 phox swift dephosphorylation (within 1 minute) in parallel with the start of p47 phox phosphorylation. p40 phox was then rephosphorylated, and this was accompanied with an increase in ROS production. Depletion of p40 phox resulted in ≈67% loss in agonist-induced ROS production despite the presence of p47 phox . These were further supported by experiments on mouse aortas stimulated with angiotensin II. Conclusion— p40 phox is prephosphorylated in resting endothelial cells and can compensate p47 phox in keeping basal ROS production. Dephosphorylation of p40 phox is a prerequisite for agonist-induced p47 phox phosphorylation, and p40 phox through its dynamic dephosphorylation and rephosphorylation is involved in the regulation of agonist-induced ROS production.
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NOX2 activation requires p22phox, p47phox, p67phox, p40phox, and Rac1.
“NOX2 activation requires p22phox, p47phox, p67phox, p40phox, and Rac1 (276).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [276]
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Cites
HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell Line
Ades, E. W., Candal, F. J., Swerlick, R. A., George, V. G., Summers, S., Bosse, D. C., & Lawley, T. J. (1992) · Journal of Investigative Dermatology · doi:10.1111/1523-1747.ep12613748 · Cited by 1,265 · see citing works
Griendling, K. K., Sorescu, D., Lassegue, B. P., & Ushio‐Fukai, M. (2000) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/01.atv.20.10.2175 · Cited by 955 · see citing works
Functional analysis of Nox4 reveals unique characteristics compared to other NADPH oxidases
Martyn, K. D., Frederick, L. M., von Loehneysen, K., Dinauer, M. C., & Knaus, U. G. (2006) · Cellular Signalling · doi:10.1016/j.cellsig.2005.03.023 · Cited by 783 · see citing works
Regulation of Nox and Duox enzymatic activity and expression.
Lambeth, J., Kawahara, T., & Diebold, B. (2007) · Free Radical Biology & Medicine · doi:10.1016/j.freeradbiomed.2007.03.028 · Cited by 509 · see citing works
Takeya, R., Ueno, N., Kami, K., Taura, M., Kohjima, M., Izaki, T., Nunoi, H., & Sumimoto, H. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m212856200 · Cited by 380 · see citing works
Li, J., Mullen, A. M., Yun, S., Wientjes, F. B., Brouns, G. Y., Thrasher, A. J., & Shah, A. M. (2002) · Circulation Research · doi:10.1161/hh0202.103615 · Cited by 312 · see citing works
Self-citationMechanism of Endothelial Cell NADPH Oxidase Activation by Angiotensin II
Li, J., & Shah, A. M. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m209793200 · Cited by 307 · see citing works
Self-citationWientjes, F. B., Hsuan, J. J., Totty, N. F., & Segal, A. W. (1993) · Biochemical Journal · doi:10.1042/bj2960557 · Cited by 301 · see citing works
Nox4 Acts as a Switch Between Differentiation and Proliferation in Preadipocytes
Schröder, K., Wandzioch, K., Helmcke, I., & Brandes, R. P. (2008) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.108.174219 · Cited by 258 · see citing works
Li, J., Fan, L. M., Christie, M. R., & Shah, A. M. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.25.6.2320-2330.2005 · Cited by 192 · see citing works
Self-citationKoshkin, V., Lotan, O., & Pick, E. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.48.30326 · Cited by 158 · see citing works
NADPH Oxidase Activity Is Independent of p47
Freeman, J. L. R., & Lambeth, J. D. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.37.22578 · Cited by 157 · see citing works
Dusi, S., Donini, M., & Rossi, F. (1996) · Biochemical Journal · doi:10.1042/bj3140409 · Cited by 143 · see citing works
Membrane Binding Mechanisms of the PX Domains of NADPH Oxidase p40 and p47
Stahelin, R. V., Burian, A., Bruzik, K. S., Murray, D., & Cho, W. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m212579200 · Cited by 143 · see citing works
Crucial role of nitric oxide synthases system in endothelium-dependent hyperpolarization in mice
Takaki, A., Morikawa, K., Tsutsui, M., Murayama, Y., Tekeş, E., Yamagishi, H., Ohashi, J., Yada, T., Yanagihara, N., & Shimokawa, H. (2008) · The Journal of Experimental Medicine · doi:10.1084/jem.20080106 · Cited by 137 · see citing works
Bouin, A., Grandvaux, N., Vignais, P. V., & Fuchs, A. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.46.30097 · Cited by 117 · see citing works
p40 Down-regulates NADPH Oxidase Activity through Interactions with Its SH3 Domain
Sathyamoorthy, M., de Mendez, I., Adams, A. G., & Leto, T. L. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.14.9141 · Cited by 112 · see citing works
Li, J., Wheatcroft, S. B., Fan, L. M., Kearney, M. T., & Shah, A. M. (2004) · Circulation · doi:10.1161/01.cir.0000118463.23388.b9 · Cited by 100 · see citing works
Self-citationYee, A. S., Paulson, E. K., McDevitt, M. A., Rieger‐Christ, K., Summerhayes, I. C., Berasi, S. P., Kim, J., Huang, C.-Y., & Zhang, X. (2004) · Gene · doi:10.1016/j.gene.2004.04.004 · Cited by 98 · see citing works
Fuchs, A., Dagher, M., & Vignais, P. V. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.11.5695 · Cited by 86 · see citing works
The NADPH Oxidase Components p47phox and p40phox Bind to Moesin through Their PX Domain
Wientjes, F. B., Reeves, E. P., Šoškić, V., Furthmayr, H., & Segal, A. W. (2001) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.2001.5982 · Cited by 82 · see citing works
HBP1 Repression of the p47phox Gene: Cell Cycle Regulation via the NADPH Oxidase
Berasi, S. P., Xiu, M., Yee, A. S., & Paulson, K. E. (2004) · Molecular and Cellular Biology · doi:10.1128/mcb.24.7.3011-3024.2004 · Cited by 73 · see citing works
p40 : The last NADPH oxidase subunit
Matute, J. D., Arias, A. A., Dinauer, M. C., & Grajales, P. J. P. (2005) · Blood Cells Molecules and Diseases · doi:10.1016/j.bcmd.2005.06.010 · Cited by 72 · see citing works
Phosphorylated p40PHOX as a Negative Regulator of NADPH Oxidase
Lopes, L. R., Dagher, M., Gutierrez, A., Young, B. M., Bouin, A., Fuchs, A., & BABIOR, B. M. (2004) · Biochemistry · doi:10.1021/bi035636s · Cited by 72 · see citing works
The HBP1 Transcriptional Repressor Participates in RAS-Induced Premature Senescence
Zhang, X., Kim, J., Ruthazer, R., McDevitt, M. A., Wazer, D. E., Paulson, K. E., & Yee, A. S. (2006) · Molecular and Cellular Biology · doi:10.1128/mcb.00604-06 · Cited by 69 · see citing works
Nox2 regulates endothelial cell cycle arrest and apoptosis via p21cip1 and p53
Li, J., Fan, L. M., George, V. T., & Brooks, G. (2007) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2007.06.001 · Cited by 65 · see citing works
Self-citationThe HMG-box transcription factor HBP1 is targeted by the pocket proteins and E1A
Lavender, P., Vandel, L., Bannister, A. J., & Kouzarides, T. (1997) · Oncogene · doi:10.1038/sj.onc.1201243 · Cited by 61 · see citing works
Phosphorylation of p40-phox during activation of neutrophil NADPH oxidase
Someya, A., Nunoi, H., Hasebe, T., & Nagaoka, I. (1999) · Journal of Leukocyte Biology · doi:10.1002/jlb.66.5.851 · Cited by 35 · see citing works
Lin, K., Zhao, W.-G., Bhatnagar, J., Zhao, W.-D., Lu, J.-P., Simko, S. J., Schueneman, A., & Austin, G. (2001) · Leukemia · doi:10.1038/sj.leu.2402071 · Cited by 27 · see citing works
Oxidant-Dependent Phosphorylation of p40phox in B Lymphocytes
Grandvaux, N., Elsen, S., & Vignais, P. V. (2001) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.2001.5665 · Cited by 16 · see citing works
Flagged first, then by citations
1 of the 31 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Biochemistry, Physiology, and Pathophysiology of NADPH Oxidases in the Cardiovascular System
Lassegue, B. P., Martín, A. S., & Griendling, K. K. (2012) · Circulation Research · doi:10.1161/circresaha.111.243972 · Cited by 768 · see citing works
Nox family NADPH oxidases: Molecular mechanisms of activation.
Brandes, R., Weissmann, N., & Schröder, K. (2014) · Free Radical Biology & Medicine · doi:10.1016/j.freeradbiomed.2014.07.046 · Cited by 660 · see citing works
Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation
Tejero, J., Shiva, S., & Gladwin, M. T. (2019) · Physiological Reviews · doi:10.1152/physrev.00036.2017 · Cited by 541 · see citing works
Molecular Biology of Atherosclerosis
Hopkins, P. N. (2013) · Physiological Reviews · doi:10.1152/physrev.00004.2012 · Cited by 456 · see citing works
Endothelial NADPH oxidases: which NOX to target in vascular disease?
Drummond, G. R., & Sobey, C. G. (2014) · Trends in Endocrinology & Metabolism · doi:10.1016/j.tem.2014.06.012 · Cited by 315 · see citing works
Cahill‐Smith, S., & Li, J. (2014) · British Journal of Clinical Pharmacology · doi:10.1111/bcp.12357 · Cited by 106 · see citing works
Self-citationEffects of angiotensin II on the cerebral circulation: role of oxidative stress
Silva, T. M. D., & Faraci, F. M. (2013) · Frontiers in Physiology · doi:10.3389/fphys.2012.00484 · Cited by 75 · see citing works
Thakur, S., Du, J., Hourani, S. M. O., Ledent, C. A., & Li, J. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.184606 · Cited by 74 · see citing works
Self-citationOrganizers and activators: Cytosolic Nox proteins impacting on vascular function
Schröder, K., Weißmann, N., & Brandes, R. P. (2017) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2017.03.017 · Cited by 72 · see citing works
Du, J., Fan, L. M., Mai, A., & Li, J. (2013) · British Journal of Pharmacology · doi:10.1111/bph.12336 · Cited by 67 · see citing works
Self-citationMeijles, D. N., Fan, L. M., Howlin, B. J., & Li, J. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.561159 · Cited by 66 · see citing works
Self-citationLin, L., Pang, W., Chen, K., Wang, F., Gengler, J., Sun, Y., & Tong, Q. (2012) · American Journal of Physiology-Endocrinology and Metabolism · doi:10.1152/ajpendo.00462.2011 · Cited by 54 · see citing works
Tanaka, L. Y., Bechara, L. R. G., dos Santos, A. M., Jordão, C. P., de Sousa, L. G. O., Bartholomeu, T., Ventura, L. I., Laurindo, F. R. M., & Ramires, P. R. (2015) · Nitric Oxide · doi:10.1016/j.niox.2015.01.003 · Cited by 50 · see citing works
Ueyama, T., Nakakita, J., Nakamura, T., Kobayashi, T., Kobayashi, T., Son, J., Sakuma, M., Sakaguchi, H., Leto, T. L., & Saito, N. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m111.237289 · Cited by 44 · see citing works
Fan, L. M., & Li, J. (2014) · Journal of Pharmacological and Toxicological Methods · doi:10.1016/j.vascn.2014.03.173 · Cited by 43 · see citing works
Self-citationDeng, B., Xie, S., Wang, J., Xia, Z., & Nie, R. (2012) · Journal of Vascular Research · doi:10.1159/000332337 · Cited by 39 · see citing works
Nox2-derived ROS in PPARγ signaling and cell-cycle progression of lung alveolar epithelial cells
Tickner, J. C., Fan, L. M., Du, J., Meijles, D. N., & Li, J. (2011) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2011.05.027 · Cited by 36 · see citing works
Self-citationMalam, Z., Parodo, J. C., Waheed, F., Szászi, K., Kapùs, A., & Marshall, J. C. (2011) · The Journal of Immunology · doi:10.4049/jimmunol.1003706 · Cited by 27 · see citing works
Teng, L., Fan, L. M., Meijles, D. N., & Li, J. (2012) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.112.247775 · Cited by 23 · see citing works
Self-citationMeijles, D. N., Fan, L. M., Ghazaly, M. M., Howlin, B. J., Krönke, M., Brooks, G., & Li, J. (2016) · Circulation · doi:10.1161/circulationaha.116.021993 · Cited by 18 · see citing works
Self-citationThe Role of PB1 Domain Proteins in Endothelial Cell Dysfunction and Disease
Burke, R. M., & Berk, B. C. (2015) · Antioxidants and Redox Signaling · doi:10.1089/ars.2014.6182 · Cited by 11 · see citing works
Ezetimibe and Reactive Oxygen Species
Yamaoka‐Tojo, M., Tojo, T., Takahira, N., Masuda, T., & Izumi, T. (2010) · Current Vascular Pharmacology · doi:10.2174/157016111793744652 · Cited by 11 · see citing works
Efficacy of silymarin in treatment of COPD via P47phox signaling pathway
Xu, L., Song, Q., Ouyang, Z., Zheng, M., Zhang, X., & Zhang, C. (2021) · Food Science and Technology · doi:10.1590/fst.52821 · Cited by 3 · see citing works
Vascular Reactive Oxygen Species Biology – Insights from Transgenic and Knockout Mouse Models
Tan, S. M., & de Haan, J. B. (2014) · Springer eBooks · doi:10.1007/978-3-642-30018-9_22
- Authors
- Fan, L. M., Teng, L., & Li, J.-M.
- Affiliations
- University of Surrey (GB)
- Venue
- Arteriosclerosis, Thrombosis, and Vascular Biology
- Published
- 2009-10
- Volume
- 29
- Issue
- 10
- Pages
- 1651-1656
- ISSN
- 1079-5642 (ISSN-L)
- 1524-4636
- PMC
- PMC2888064
- OpenAlex
- W2102340898
- Cited by
- 24
- References
- 31
- Citations by year
- 2021: 1 · 2018: 1 · 2017: 1 · 2016: 1 · 2015: 2 · 2014: 6 · 2013: 3 · 2012: 4
- FWCI
- 1.99
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Arteriosclerosis Thrombosis and Vascular Biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Surrey Research Insight Open Access (The University of Surrey) (submittedVersion)
- Open Research (University of Surrey) (submittedVersion)
- Funders
- Wellcome Trust (078637, 07863/Z/05/Z)
- Biotechnology and Biological Sciences Research Council (BB/D009510/1)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- p40phox, p47phox, NADPH oxidase, reactive oxygen species, microvascular endothelial cells, HBP1, protein phosphorylation, dephosphorylation, angiotensin II, coronary endothelium
- MeSH terms
- Angiotensin II, Animals, Cells, Cultured, Coronary Vessels, High Mobility Group Proteins, Microcirculation, Phosphoproteins, Phosphorylation, Repressor Proteins, RNA, Messenger, Tetradecanoylphorbol Acetate, Reactive Oxygen Species, Mice, Knockout, NADPH Oxidases, Endothelial Cells, Cell Proliferation, Mice
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