Abstract
PX domains are found in a variety of proteins that associate with cell membranes, but their molecular function has remained obscure. We show here that the PX domains in p47phox and p40phox subunits of the phagocyte NADPH oxidase bind to phosphatidylinositol-3,4-bisphosphate (PtdIns(3,4)P2) and phosphatidylinositol-3-phosphate (PtdIns(3)P), respectively. We also show that an Arg-to-Gln mutation in the PX domain of p47phox, which is found in patients with chronic granulomatous disease, eliminates phosphoinositide binding, as does the analogous mutation in the PX domain of p40phox. The PX domain of p40phox localizes specifically to PtdIns(3)P-enriched early endosomes, and this localization is disrupted by inhibition of phosphoinositide-3-OH kinase (PI(3)K) or by the Arg-to-Gln point mutation. These findings provide a molecular foundation to understand the role of PI(3)K in regulating neutrophil function and inflammation, and to identify PX domains as specific phosphoinositide-binding modules involved in signal transduction events in eukaryotic cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ponting, C. P. Protein Sci. 5, 2353–2537 (1996).
Kurten, R. C., Cadena, D. L. & Gill, G. N. Science 272, 1008–1010 (1996).
Haft, C. R., de la Luz Sierra, M., Barr, V. A., Haft, D. H. & Taylor, S. I. Mol. Cell Biol. 18, 7278–7287 (1998).
Phillips, S A, Barr, V A, Haft, D H, Taylor, S I & Haft, C R. J. Biol. Chem. 276, 5074–5084 (2001).
Horazdovsky, B. F. et al. Mol. Biol. Cell 8, 1529–1541 (1997).
Sato, T. K., Darsow, T. & Emr, S. D. Mol. Cell Biol. 18, 5308–5319 (1998).
Ekena, K. & Stevens, T. H. Mol. Cell Biol. 15, 1671–1678 (1995).
Liu, D., Yang, X. & Songyang, Z. Curr. Biol. 10, 1233–1236 (2000).
Babior, B. M. Blood 93, 1464–1476 (1999).
Nauseef, W. M. Proc. Assoc. Am. Physicians 111, 373–382 (1999).
Wientjes, F. B., Segal, A. W. & Hartwig, J. H. J. Leukoc. Biol. 61, 303–312 (1997).
Goldblatt, D. & Thrasher, A. J. Clin. Exp. Immunol. 122, 1–9 (2000).
Stephens, L., Jackson, T. & Hawkins, P. T. J. Biol. Chem. 268, 17162–17172 (1993).
Arcaro, A. & Wymann, M. P. Biochem. J. 296, 297–301 (1993).
Okada, T., Sakuma, L., Fukui, Y., Hazeki, O. & Ui, M. J. Biol. Chem. 269, 3563–3567 (1994).
Hirsch, E. et al. Science 287, 1049–1053 (2000).
Yaffe, M. B. & Smerdon, S. J. Structure 9, R33–R38 (2001).
Fruman, D. A., Rameh, L. E. & Cantley, L. C. Cell 97, 817–820 (1999).
Dowler, S. et al. Biochem. J. 351, 19–31 (2000).
Klarlund, J. K. et al. Science 275, 1927–1930 (1997).
Kavran, J. M. et al. J. Biol. Chem. 273, 30497–30508 (1998).
Noack, D. et al. Blood 97, 305–311 (2001).
Patki, V. et al. Proc. Natl Acad. Sci. USA 94, 7326–7330 (1997).
Lawe, D. C., Patki, V., Heller-Harrison, R., Lambright, D. & Corvera, S. J. Biol. Chem. 275, 3699–3705 (2000).
Gillooly, D. J. et al. EMBO J. 19, 4577–4588 (2000).
Patki, V., Lawe, D. C., Corvera, S., Virbasius, J. V. & Chawla, A. Nature 394, 433–434 (1998).
Stenmark, H., Aasland, R., Toh, B. H. & D'Arrigo, A. J. Biol. Chem. 271, 24048–24054 (1996).
Ford, M. G. et al. Science 291, 1051–1055 (2001).
Itoh, T. et al. Science 291, 1047–1051 (2001).
Mao, Y., Chen, J., Maynard, J. A., Zhang, B. & Quiocho, F. A. Cell 104, 433–440 (2001).
Botelho, R. J. et al. J. Cell Biol. 151, 1353–1368 (2000).
Acknowledgements
We thank J. Miyagi for technical assistance, S.-Y. Pai for suggesting the R42Q mutation, and members of M.B.Y.'s laboratory for discussions. F.K. was supported by fellowships from the Cell Science Research Foundation and Sankyo Foundation of Life Science. S.J.F. was supported by a Howard Hughes Medical Institute Postdoctoral Fellowship. This work was funded by NIH grants to M.B.Y. and L.C.C, and a Burroughs-Wellcome Career Development Award to M.B.Y.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kanai, F., Liu, H., Field, S. et al. The PX domains of p47phox and p40phox bind to lipid products of PI(3)K. Nat Cell Biol 3, 675–678 (2001). https://doi.org/10.1038/35083070
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/35083070
This article is cited by
-
Unexpected inhibition of the lipid kinase PIKfyve reveals an epistatic role for p38 MAPKs in endolysosomal fission and volume control
Cell Death & Disease (2024)
-
Hippo pathway regulation by phosphatidylinositol transfer protein and phosphoinositides
Nature Chemical Biology (2022)
-
Characterization of the binding of cytosolic phospholipase A2 alpha and NOX2 NADPH oxidase in mouse macrophages
Molecular Biology Reports (2022)
-
Redefining the specificity of phosphoinositide-binding by human PH domain-containing proteins
Nature Communications (2021)
-
Compartmentalized replication organelle of flavivirus at the ER and the factors involved
Cellular and Molecular Life Sciences (2021)