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(The FASEB Journal. 2008;22:lb552)
© 2008 FASEB
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lb552

NADPH oxidase flavocytochrome b localizes to Rab11-positive recycling endosomes in macrophages

Amy Jo Casbon1, Kenneth W. Dunn2, Lee-Ann Allen3 and Mary C. Dinauer1

1 Dept of Pediatrics, Wells Ctr Pediatric Res, Dept of Micro & Immunology,
2 Div of Neph, Indiana Univ Sch of Med, Indianapolis, IN,
3 Dept of Med & Microbiology, VA Med Ctr, Univ of Iowa, Iowa City, IA

ABSTRACT

Altering the localization of flavocytochrome b, the redox center of the phagocyte NADPH oxidase, has been suggested to be a mechanism by which pathogens evade oxidative killing in macrophages. Flavocytochrome b is a membrane heterodimer consisting of a large subunit gp91phox (NOX2) and a smaller subunit, p22phox. While flavocytochrome b is found primarily in specific granules in neutrophils, the localization of flavocytochrome b in macrophages has not been well characterized. The goal of these studies was to determine the subcellular localization of flavocytochrome b in CHO-K1 (Chinese Hamster Ovary), RAW264.7 (murine macrophage), and primary murine bone marrow derived macrophages utilizing live imaging of fluorescent-tagged gp91phox and p22phox subunits and immunofluorescent staining. Results were similar in all three cell types. We found that unassembled subunits localize to the endoplasmic reticulum, while the flavocytochrome b heterodimer efficiently localizes not only to the plasma membrane, but also to Rab11-positive recycling endosomes. These studies show for the first time that flavocytochrome b localizes to an intracellular compartment in macrophages that recycles to the plasma membrane. Future studies will address whether flavocytochrome b localization to recycling endosomes is disrupted by pathogens known to evade oxidative killing.

This work was supported in part by National Institutes of Health Grants RO1 HL45635 (to M.C.D) and T32-DK007519 (to A.J.C).





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