NDLI: A Fluorescently Tagged C-Terminal Fragment of p47 phox Detects NADPH Oxidase Dynamics during Phagocytosis

Content Provider	PubMed Central
Author	Li, Xing Jun Tian, Wei Stull, Natalie D. Grinstein, Sergio Atkinson, Simon Dinauer, Mary C.
Editor	Lippincott-schwartz, Jennifer
Copyright Year	2009
Abstract	The assembly of cytosolic p47 phox and p67 phox with flavocytochrome b 558 at the membrane is crucial for activating the leukocyte NADPH oxidase that generates superoxide for microbial killing. p47 phox and p67 phox are linked via a high-affinity, tail-to-tail interaction involving a proline-rich region (PRR) and a C-terminal SH3 domain (SH3b), respectively, in their C-termini. This interaction mediates p67 phox translocation in neutrophils, but is not required for oxidase activity in model systems. Here we examined phagocytosis-induced NADPH oxidase assembly, showing the sequential recruitment of YFP-tagged p67 phox to the phagosomal cup, and, after phagosome internalization, a probe for PI(3)P followed by a YFP-tagged fragment derived from the p47 phox PRR. This fragment was recruited in a flavocytochrome b 558-dependent, p67 phox -specific, and PI(3)P-independent manner. These findings indicate that p47PRR fragment probes the status of the p67 phox SH3b domain and suggest that the p47 phox /p67 phox tail-to-tail interaction is disrupted after oxidase assembly such that the p67 phox -SH3b domain becomes accessible. Superoxide generation was sustained within phagosomes, indicating that this change does not correlate with loss of enzyme activity. This study defines a sequence of events during phagocytosis-induced NADPH oxidase assembly and provides experimental evidence that intermolecular interactions within this complex are dynamic and modulated after assembly on phagosomes.
Related Links	http://dx.doi.org/10.1091/mbc.e08-06-0620
Ending Page	1532
Page Count	13
Starting Page	1520
File Format	PDF
ISSN	10591524
e-ISSN	19394586
Journal	Molecular Biology of the Cell
Issue Number	5
Volume Number	20
Language	English
Publisher	The American Society for Cell Biology
Publisher Date	2009-01-01
Access Restriction	Open
Rights Holder	The American Society for Cell Biology
Subject Keyword	Cell Biology Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Molecular Biology

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