The JI
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     
 

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Taylor, R. M.
Right arrow Articles by Jesaitis, A. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Taylor, R. M.
Right arrow Articles by Jesaitis, A. J.
The Journal of Immunology, 2004, 173: 7349-7357.
Copyright © 2004 by The American Association of Immunologists

Site-Specific Inhibitors of NADPH Oxidase Activity and Structural Probes of Flavocytochrome b: Characterization of Six Monoclonal Antibodies to the p22phox Subunit1

Ross M. Taylor*, James B. Burritt*, Danas Baniulis*, Thomas R. Foubert*, Connie I. Lord*, Mary C. Dinauer{dagger}, Charles A. Parkos{ddagger} and Algirdas J. Jesaitis2,*

* Department of Microbiology, Montana State University, Bozeman, Montana 59717; {dagger} Departments of Pediatrics and Medical and Molecular Genetics, Indiana University Medical Center, Indianapolis, IN 46202; and {ddagger} Department of Pathology and Laboratory Medicine, Division of Gastrointestinal Pathology, Emory University, Atlanta, GA 30322

The integral membrane protein flavocytochrome b (Cyt b) is the catalytic core of the human phagocyte NADPH oxidase, an enzyme complex that initiates a cascade of reactive oxygen species important in the elimination of infectious agents. This study reports the generation and characterization of six mAbs (NS1, NS2, NS5, CS6, CS8, and CS9) that recognize the p22phox subunit of the Cyt b heterodimer. Each of the mAbs specifically detected p22phox by Western blot analysis but did not react with intact neutrophils in FACS studies. Phage display mapping identified core epitope regions recognized by mAbs NS2, NS5, CS6, CS8, and CS9. Fluorescence resonance energy transfer experiments indicated that mAbs CS6 and CS8 efficiently compete with Cascade Blue-labeled mAb 44.1 (a previously characterized, p22phox-specific mAb) for binding to Cyt b, supporting phage display results suggesting that all three Abs recognize a common region of p22phox. Energy transfer experiments also suggested the spatial proximity of the mAb CS9 and mAb NS1 binding sites to the mAb 44.1 epitope, while indicating a more distant proximity between the mAb NS5 and mAb 44.1 epitopes. Cell-free oxidase assays demonstrated the ability of mAb CS9 to markedly inhibit superoxide production in a concentration-dependent manner, with more moderate levels of inhibition observed for mAbs NS1, NS5, CS6, and CS8. A combination of computational predictions, available experimental data, and results obtained with the mAbs reported in this study was used to generate a novel topology model of p22phox.




This article has been cited by other articles:


Home page
 J. Leukoc. Biol.Home page
M. Mori, G. Li, M. Hashimoto, A. Nishio, H. Tomozawa, N. Suzuki, S.-i. Usami, K. Higuchi, and K. Matsumoto
Pivotal Advance: Eosinophilia in the MES rat strain is caused by a loss-of-function mutation in the gene for cytochrome b(-245), alpha polypeptide (Cyba)
J. Leukoc. Biol., September 1, 2009; 86(3): 473 - 478.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
A.-J. Casbon, L.-A. H. Allen, K. W. Dunn, and M. C. Dinauer
Macrophage NADPH Oxidase Flavocytochrome b Localizes to the Plasma Membrane and Rab11-Positive Recycling Endosomes
J. Immunol., February 15, 2009; 182(4): 2325 - 2339.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. von Lohneysen, D. Noack, A. J. Jesaitis, M. C. Dinauer, and U. G. Knaus
Mutational Analysis Reveals Distinct Features of the Nox4-p22phox Complex
J. Biol. Chem., December 12, 2008; 283(50): 35273 - 35282.
[Abstract] [Full Text] [PDF]

Home page
 NEJMHome page
J. B. Harris, I. C. Michelow, S. J. Westra, and R. L. Kradin
Case 21-2008 -- An 11-Month-Old Boy with Fever and Pulmonary Infiltrates
N. Engl. J. Med., July 10, 2008; 359(2): 178 - 187.
[Full Text] [PDF]

Home page
 Physiol. Rev.Home page
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]

Home page
 J. Biol. Chem.Home page
R. M. Taylor, D. Baniulis, J. B. Burritt, J. M. Gripentrog, C. I. Lord, M. H. Riesselman, W. S. Maaty, B. P. Bothner, T. E. Angel, E. A. Dratz, et al.
Analysis of Human Phagocyte Flavocytochrome b558 by Mass Spectrometry
J. Biol. Chem., December 1, 2006; 281(48): 37045 - 37056.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Zhu, C. C. Marchal, A.-J. Casbon, N. Stull, K. von Lohneysen, U. G. Knaus, A. J. Jesaitis, S. McCormick, W. M. Nauseef, and M. C. Dinauer
Deletion Mutagenesis of p22phox Subunit of Flavocytochrome b558: IDENTIFICATION OF REGIONS CRITICAL FOR gp91phox MATURATION AND NADPH OXIDASE ACTIVITY
J. Biol. Chem., October 13, 2006; 281(41): 30336 - 30346.
[Abstract] [Full Text] [PDF]

Home page
 NEJMHome page
D. W. Moskowitz, S. Khan, H. K. Eltzschig, J. Karhausen, V. A.J. Kempf, W. M. Nauseef, E. K. Weir, J. Lopez-Barneo, and S. L. Archer
Acute oxygen-sensing mechanisms.
N. Engl. J. Med., March 2, 2006; 354(9): 975 - 977.
[Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Website Copyright © 2004 by The American Association of Immunologists, Inc. All rights reserved.
All Contents Copyright © 2004 by The American Association of Immunologists, Inc. All rights reserved.