Macrophage NADPH oxidase flavocytochrome b localizes to the plasma membrane and Rab11-positive recycling endosomes

Amy Jo Casbon, Lee Ann H Allen, Kenneth Dunn, Mary C. Dinauer

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Flavocytochrome b558, the catalytic core of the phagocytic NADPH oxidase, mediates the transfer of electrons from NADPH to molecular oxygen to generate superoxide for host defense. Flavocytochrome b is a membrane heterodimer consisting of a large subunit gp91phox (NOX2) and a smaller subunit, p22phox. Although in neutrophils flavocytochrome b has been shown to localize to the plasma membrane and specific granules, little is known about its distribution in macrophages. Using immunofluorescent staining and live cell imaging of fluorescently tagged gp91phox and p22 phox, we demonstrate in a Chinese hamster ovary cell model system and in RAW 264.7 and primary murine bone marrow-derived macrophages that flavocytochrome b is found in the Rab11-positive recycling endocytic compartment, as well as in Rab5-positive early endosomes and plasma membrane. Additionally, we show that unassembled p22phox and gp91 phox subunits localize to the endoplasmic reticulum, which redistribute to the cell surface and endosomal compartments following heterodimer formation. These studies show for the first time that flavocytochrome b localizes to intracellular compartments in macrophages that recycle to the plasma membrane, which may act as a reservoir to deliver flavocytochrome b to the cell surface and phagosome membranes.

Original languageEnglish
Pages (from-to)2325-2339
Number of pages15
JournalJournal of Immunology
Volume182
Issue number4
DOIs
StatePublished - Feb 15 2009

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NADPH Oxidase
Endosomes
Macrophages
Cell Membrane
Phagosomes
Membranes
Recycling
Cricetulus
NADP
Superoxides
Endoplasmic Reticulum
cytochrome b558
Ovary
Catalytic Domain
Neutrophils
Electrons
Staining and Labeling
Oxygen

ASJC Scopus subject areas

  • Immunology
  • Medicine(all)

Cite this

Macrophage NADPH oxidase flavocytochrome b localizes to the plasma membrane and Rab11-positive recycling endosomes. / Casbon, Amy Jo; Allen, Lee Ann H; Dunn, Kenneth; Dinauer, Mary C.

In: Journal of Immunology, Vol. 182, No. 4, 15.02.2009, p. 2325-2339.

Research output: Contribution to journalArticle

Casbon, Amy Jo ; Allen, Lee Ann H ; Dunn, Kenneth ; Dinauer, Mary C. / Macrophage NADPH oxidase flavocytochrome b localizes to the plasma membrane and Rab11-positive recycling endosomes. In: Journal of Immunology. 2009 ; Vol. 182, No. 4. pp. 2325-2339.
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